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340
SECTION 11.2 Metabolism of Ammonia CHAPTER 17Protein and Amino Acid 341
Metabolism
Brain takes up significant quantities of valine derived from intestinal metabolism (e.g., glutamine) and
and may be a major (if not primary) site of ingested protein. The ammonia diffuses across the
utilization of branched-chain amino acids. intestinal mucosa to the portal blood and is converted
Glutamate, aspartate, and glycine are to urea in the liver.
neurotransmitters. Glutamate is a precursor of y-
aminobutyrate; tyrosine of dopamine, norepinephrine,
and epinephrine; and tryptophan of serotonin, all of
which are neurotransmitters. Inactivation of
neurotransmitters involves deamination with
production of ammonia, which is removed by formation
of glutamine. N-acetylaspartate occurs in high levels in
the brain but its function is not known. It is synthesized
from acetyl-CoA and aspartic acid catalyzed by
acetyl-CoA aspartate N-acetyl transferase. As-
partoacylase catalyzes the hydrolysis of N-
acetylaspartate to acetate and aspartic acid. The
deficiency of aspartoa- cylase, which is inherited as an
autosomal recessive trait, is associated with
degenerative brain changes. Patients of this disorder,
also known as Canavan dystrophy, are usually of
Eastern European Jewish heritage.
17.2 Metabolism of Ammonia
Ammonia (at physiological pH, 98.5% exists as NHt),

the highly toxic product of protein catabolism, is
rapidly inactivated by a variety of reactions. Some
products of these reactions are utilized for other
purposes (thus salvaging a portion of the amino
nitrogen), while others are excreted. The excreted form
varies quite widely among vertebrate and invertebrate
animals. The development of a pathway for nitrogen
disposal in a species appears to depend chiefly on the
availability of water. Thus, urea is excreted in
terrestrial vertebrates (ureotelic organisms); ammonia
in aquatic animals (ammonotelic organisms); and uric
acid (in semisolid form) in birds and land-dwelling
reptiles (uricotelic organisms). During their aquatic
phase of development amphibia excrete ammonia but
the adult frog excretes urea; during metamorphosis the
liver produces the enzymes required for their synthesis. In
humans, ammonia is excreted mostly as urea, which is
highly water-soluble, is distributed throughout
extracellular and intracellular body water, is nontoxic and
metabolically inert, has a high nitrogen content (47%),
and is excreted via the kidneys.
Ammonia is produced by deamination of glutamine,
glutamate, other amino acids, and adenylate. A consid-
erable quantity is derived from intestinal bacterial en-
zymes acting on urea and other nitrogenous compounds.
The urea comes from body fluids that diffuse into the in-
testine, and the other nitrogenous products are
341
SECTION 11.2 Metabolism of Ammonia CHAPTER 17 Protein and Amino Acid 341
Metabolism
is not converted to urea may be incorporated into
Ammonia is particularly toxic to brain but not
to other tissues, even though levels in those tissues glutamine catalyzed
may increase under normal physiological conditions
(e.g., in muscle during heavy exercise in kidney during
metabolic acidosis). Several hypotheses have been
suggested to explain the mechanism of neurotoxicity.
In brain mitochondria, excess ammonia may drive
the reductive amination of o-ketoglutarate by
glutamate dehydrogenase. This step may deplete a
key intermediate of the TCA cycle and lead to its
impairment, with severe inhibition of respiration and
considerable stimulation of glycolysis. Since the
[NAD+J/[NADHl ratio will be high in mitochondria,
there will be a decrease in the rate of production of
ATP. This hypothesis does not explain why the same
result does not occur in tissues that are not
affected by ammonia. A more plau- sible hypothesis
is depletion of glutamate which is an excitatory
neurotransmitter. Glutamine, synthesized and stored in
glial cells, is the most likely precursor of glutamate.
It is transported into the neurons and hydrolyzed by
glutaminase. Ammonia inhibits glutaminase and de-
pletes the glutamate concentration. A third
hypothesis invokes neuronal membrane dysfunction,
since elevated levels of ammonia produce increased
permeability to K+ and c1- ions, while glycolysis
increases H" ion concentration (NHt stimulates 6-
phosphofructokinase; Chap- ter 13). Encephalopathy
of hyperammonemia is characterized by brain edema
and astrocyte swelling. Edema and swelling have been
attributed to intracellular accumulation of glutamine
which causes osmotic shifts of water into the cell.
Behavioral disorders such as anorexia, sleep
distur-
bances, and pain insensitivity associated with
hyperammonemia have been attributed to increased
tryptophan transport across the blood-brain barrier and
the accumulation of its metabolites. Two of the
tryptophan-derived metabolites are serotonin and
quinolinic acid (discussed later). The latter is an
excitotoxin at the N-methyl-D- aspartate (NMDA)
glutamate receptors. Thus, the mechanism of the
ammonium-induced neurological abnormalities is
multifactorial. Normally only small amounts ofNH3 (i.e.,
NHt) are present in plasma, since NH3 is rapidly re-
moved by reactions in tissues of glutamate dehydrogenase,
glutamine synthase, and urea formation.
Urea Synthesis
Ammonia contained in the blood flowing through the
hepatic lobule is removed by the hepatocytes and
converted into urea. Periportal hepatocytes are the
predominant sites of urea formation. Any ammonia that
342
Metabolism
NH; 2ATP 2ADP +

P,
\, ) Carbamoyl�P,
c>o, CPSI pbcspnate ( \
N-
4 Ornithine
Acetylglutamate
CoASH--f Citrulline
JNAGS
Acetyl-CoA + Glutamate
Mitochondrial inner
membrane Mitochondrial
outer membrane
Urearrnithine Citrulr
ine
A ATP Aspartate
H,O AS
AMP +PP,
. .V'""'""
Fumarate
FIGURE 17-7
Formation of urea in hepatocytes. NAGS = N-acetylglutamate synthase; CPS! = carbamoylphosphate synthase I;
OCT= omithine carbamoyltransferase; C-OT = citrulline-ornithine translocase; AS = argininosuccinate synthase;
AL= argininosuccinate lyase; A= arginase.--©--+ indicates the absolute requirement of N-acetylglutamate for
CPS! activity.
by glutamine synthase located in pericentral hepatocytes. of CoA derivatives that are also competitive inhibitors
Formation of urea requires the combined action of two of N-acetylglutamate synthase and inhibitors of CPSI.
enzymes to produce carbamoyl phosphate and of four Hyperammonemia often accompanies organic
enzymes that function in a cyclic manner in the urea acidemias.
cycle (Figure 17-7). Although some of these enzymes
occur in extrahepatic tissues and urea formation has been coo- coo-
l Arg I
shown to occur in several cell lines in tissue culture,
0 (CH2lz (CH2lz
the most important physiological site of urea formation II I :EB I
is the liver. In hepatocytes the first three enzymes are CH3C-SCoA + CHNH3+ -f- CoASH + CHNHCOCH3 + H +
mitochondrial and the others are cytosolic. A citrulline- I \ I
coo- ', ', e
coo-
'
ornithine antiport is located in the inner mitochondrial Acetyl-CoA Glutamate ' - - - - N-Acetyl-
membrane. glutamate
acids produced in excess compete for CoA-SH for

Formation of Carbamoyl Phosphate formation
Carbamoyl phosphate synthesis requires amino acid
acetyltransferase (N-acetylglutamate synthase, mitochon-
drial) and carbamoyl-phosphate synthase I (CPSI). N-
Acetylglutamate (NAG) is an obligatory positive effector
of CPSI. NAG synthase is under positive allosteric mod-
ulation by arginine and product inhibition by NAG. De-
pletion of CoA-SH decreases NAG synthesis and ureage-
nesis. This situation can occur in organic acidemias
(e.g., propionic acidemia; Chapter 18), in which organic
343
Metabolism
CPSI catalyzes the reaction
NAG.K'.Mg2'
NH,++ HC0 + 2ATP4 3- -
0
II
H2N-C-OPO/- + 2ADP3- + p/- + 2H +
NAG binding changes the conformation and subunit

structure of CPSI, with preponderance of the monomers.
Carbamoylglutamate is also an activator of CPSI. Glu-
tamate and o-ketoglutarate compete with NAG for bind-
ing. CPSI is subject to product inhibition by Mg-ADP. It
342
SECTION 11.2 Metabolism of Ammonia CHAPTER 11 343
Protein and Amino Acid Metabolism
possesses two binding sites for ATP. One ATP is utilized is transported out of the mitochondria by the citrulline-
in activation of bicarbonate by forming an enzyme-bound omithine antiporter.
carboxyphosphate that reacts with ammonium ion to form
an enzyme-bound carbamate, with elimination of inor-
ganic phosphate. Carbamoyl phosphate is generated when
the second ATP reacts with the enzyme-bound carbamate, Formation ofArgininosuccinate
with release of ADP and free enzyme. The condensation of citrulline and aspartate to argini-
In humans, there are two immunologically distinct car- nosuccinate is catalyzed by argininosuccinate synthase in
bamoyl phosphate synthases, one mitochondrial (CPSI) the cytosol and occurs in two steps. In the initial step,
and the other cytosolic (CPSII). CPSI is involved in ure- the ureido group is activated by ATP to form the enzyme-
agenesis, uses NH3 exclusively as the nitrogen donor, bound intermediate adenylylcitrulline. In the second step,
and requires binding of NAG for activity. CPSII uses nucleophilic attack of the amino group of aspartate
glutamine as substrate, is not dependent on NAG for displaces AMP and yields argininosuccinate. The overall
activity, and is required for synthesis of pyrimidine reaction is shown below:
NH2 NH COO-
I i I
C=O C=NH•-CH
I I I
NH coo- NH CH2
I l I I
CH2 CH2 COO-
CHNH3 +
I +I I +I ATP'- i + AMP 2" + PP,3" + H+
CH2 CH2
I .:::=
CH2 CH2
I
CH2
I I I
CHNH3 + coo- CHNH3 +
I I
coo- coo-
L-Citrulline L-Argininosuccinate
L-Aspartate
(Chapter 27). Normally, the mitochondrial membrane is The reaction is driven forward by hydrolysis of pyrophos-
not permeable to carbamoyl phosphate, but when the phate to inorganic phosphate. Argininosuccinate forma-
concentration increases, carbamoyl phosphate spills into tion is considered as the rate-limiting step for urea synthe-
the cytosol and promotes synthesis of orotic acid and sis. This reaction incorporates the second nitrogen atom
uridine 5' -phosphate. of the urea molecule donated by aspartate.
Formation of Citrulline Formation ofArginine and Fumarate

Ornithine carbamoyltransferase (ornithine transcar- Argininosuccinate lyase in cytosol catalyzes cleavage
bamoylase) catalyzes the condensation between car- of argininosuccinate to arginine and fumarate:
bamoyl phosphate and ornithine to yield citrulline in
mitochondria: 0
II
NH 2 co· NH2
NH2} Transferred I I I
C=NH•-cH C=NH2•
I carbamoyl
NH3 + C=o group I I I
NH CH2 NH coo-
I I I I I
CH2 NH CH2 coo- CH2 l
I I + CH
CH2 l I
O CH2 CH2 II
CH2
II
H2N-C-O-P032 - + CH2
I
I
= I
CH2
I
+P;2- + H+ I
CH2
I
CH2
HC
I
coo-
I I
Carbamoyl CHNH3 • CH2 CHNH +
CHNH3 + 3
phosphate I I I I
coo- CHNH3 +
coo- coo-
I L·Argininosuccinic acid L-Arginine Fumarate
coo-
L-Ornithine L-Citrulline
This is the pathway for synthesis of arginine, a nonessen-
Although the equilibrium constant strongly favors cit- tial amino acid; however, in the event of physiological
rulline formation, the reaction is reversible. Citrulline deficiency, as in premature infants, or a defect in any of
343
SECTION 11.2 Metabolism of Ammonia CHAPTER 11 343
the enzymes discussed above, an exogenous supply of Hyperammonemias

arginine is required. Hyperammonemias are caused by inborn errors of
ureagenesis and organic acidemias, liver immaturity
Formation of Urea and (tran- sient hyperammonemia of the newborn), and liver
Ornithine failure (hepatic encephalopathy). Neonatal
This irreversible reaction is catalyzed by arginase in the hyperammonemias are characterized by vomiting,
cytosol: lethargy, lack of appetite, seizures, and coma. The
underlying defects can be identified by appropriate
NH2 laboratory measurements (e.g., assessment of
I metabolic acidosis if present and character- ization of
C=NH2+ organic acids, urea cycle intermediates, and glycine).
I Inborn errors of the six enzymes of ureagenesis and
NH
I NH2 NAG synthase have been described. The inheritance pat-
CH2 I tern of the last is not known, but five of the urea cycle
I + H20--+C=O + defects are autosomal recessive and ornithine carbamoyl-
CH2 I
I NH2 transferase (OCT) deficiency is X-linked.
CH2 Carriers of OCT deficiency (estimated to be several
I thousand women in the U.S.A.) can be identified by admin-
CHNH3 +
I istration of a single oral dose of allopurinol, a purine ana-
coo-
logue, followed by measurement of urinary orotidine ex-
l-Arginine Urea L-
Ornithine cretion. The underlying principle of this assay is that when
the intramitochondrial carbamoyl phosphate accumulates
The urea so formed is distributed throughout the body wa-
in OCT heterozygotes, it diffuses into the cytoplasm stimu-
ter and excreted. The renal clearance of urea is less than
lating the biosynthesis of pyrimidines. One of the interme-
the glomerular filtration rate because of passive tubular
diates in this pathway-orotidine-accumulates, leading
back-diffusion. Diffusion of urea in the intestine leads to
formation of ammonia, which enters the portal blood and to orotidinuria (Figure 17-8).
is converted to urea in liver. Reentry of ornithine into mi- The sensitivity of this test is increased by increas-
tochondria initiates the next revolution of the urea cycle. ing the flux in the pyrimidine biosynthetic pathway. The
Ornithine can be converted to glutamate-y-semialdehyde enhanced flux is accomplished by allopurinol, which by
(which is in equilibrium with its cyclic form S-pyrroline- way of oxypurinol ribonucleotide inhibits the fonna-
5-carboxylate) by ornithine aminotransferase and de- tion of final product uridine 5' -phosphate (UMP) in the
carboxylated to putrescine by ornithine decarboxylase. pyrimidine biosynthesis (Chapter 27).
Ornithine is also produced in the arginine-glycine trans- Antenatal diagnosis for fetuses at risk for the urea
amidinase reaction. cycle enzyme disorders can be made by appropri-
The availability of substrates (ammonia and amino ate enzyme assays and DNA analysis in the cultured
acids) in the liver determines the amount of urea syn- amniocytes.
thesized. Urea excretion increases with increased protein Acute neonatal hyperammonemia, irrespective of
intake and decreases with decreased protein intake. cause, is a medical emergency and requires immediate
and rapid lowering of ammonia levels to prevent serious
effects on the brain. Useful measures include hemodialy-
Energetics of Ureagenesis sis, exchange transfusion, peritoneal dialysis, and admin-
The overall reaction of ureagenesis is istration of arginine hydrochloride. The general goals of
management are to
NH3 + HC03 + aspartate + 3ATP --+
urea + fumarate + 2ADP + 4Pi + AMP 1. Decrease nitrogen intake so as to minimize the
requirement for nitrogen disposal,
Hydrolysis of four high-energy phosphate groups is 2. Supplement arginine intake, and
required for the formation of one molecule of urea. If 3. Promote nitrogen excretion in forms other than urea.
fumarate is converted to aspartate (by way of malate and
oxaloacetate), one NADH molecule is generated that can The first can be accomplished by restriction of dietary
give rise to three ATP molecules through the electron protein and administration of a-keto analogues of essen-
transport chain, so that the energy expenditure becomes tial amino acids. Arginine supplementation as a precursor
one ATP molecule per each molecule of urea. of ornithine is essential to the urea cycle. The diversion
344 17.3
SECTION Metabolism of Some Individual Amino Acids CHAPTER 11Protein and Amino Acid 345
Metabolism
Acetyl-CoA NAGS
(Glutamate
MITOCHONDAION
N-Acetylglutamate
�
NH: } CPS I OTC . .
2ATP_ - Carbamoyl Phosphate -------,-++
Cttrulllne
HC03 Orni hine ·
Urea
P, Hp
Glutamine} CPS 11 'f -�D_.,_f_
_.
HC03
2ATP
- Carbamoyl Phosphate 4---1.
Aspartate
Ca rbamoyI Aspa rt ate -
Dihydro-
Orotate
NAD+
DH
NADH+W
OPAT f
r
Orotate
PAPP
PP,
. XO O . . PAT Oxipurinol
Allopu nnol- xlpurinol �Aibonucleotide
_;_. ,
PAPP pp
4-
Orotidine 5 · -Monophosphate (OMP) - - - - ._ Orotidine
OMP
Decarboxylase
I Uridine 5 ·-Monophosphate
FIGURE 17-8
The metabolic interrelationship between mitochondrial carbamoyl phosphate synthesis to urea formation and to
cytosolic carbamoyl phosphate channeled into pyrimidine biosynthesis. In ornithine lranscarbamoylase (OTC)
deficiency, mitochondrial carbamoyl phosphate diffuses into the cytosol and stimulates pyrimidine biosynthesis, leading
to orotidinuria. Administration of allopurinol augments orotidinuria by increasing the Hux in the pyrimidine
biosynthetic pathway. CPS = Carbamoyl phosphate synthase. AT= aspartate transcarbamoylase, D = dihydroorotase,
DH = dihydroorotate dehydrogenase, OPRT = orotate phosphoribosyltransferase, XO= xanthine oxidase,
PRT = phosphoribosyltransferase, PRPP = 5-phosphoribosyl-1-pyrophosphate.
of nitrogen to products other than urea is achieved by ad- phosphate, catalyzed by mitochondrial glycine synthase
ministration of sodium benzoate or sodium (or calcium) (glycine cleavage enzyme).
phenylacetate. Administration of benzoate leads to elimi- Phenylacetate or phenylbutyrate administration m-
nation of hippurate (benzoylglycine): creases excretion of phenylacetylglutamine:
0
0-
II
O
Activating onzyme
0 CH,-coo- +ATP'-+ CoASH -��- CH3-C-SCoA +AMP'-+ PP;'-
0-C-o-
11 ..
+ ATP' - + CoASH ""'""''"g °"'""° Phenylacetyl·CoA
Benzoate
CONH,
CONH, I
0 (CH,l,
I
0
II
0
I
( CH2h Conjugating enzyme
0- 3
C-N-CH
I I
+CoASH +H'
O-!-s-coA + AMP2- + pp,3- O CH3-C-SCoA + H3 •N-CH

I
coo-
CH -
H I
coo-
Benzoyl-CoA Glutamine
Phenylacetylglutamlne
The excretion of phenylacetylglutamine produces loss of

Hippurate is rapidly secreted since its clearance is five two nitrogen atoms.
times greater than its glomerular filtration rate. The glycine NAG synthase deficiency cannot be treated by admin-
nitrogen is derived from ammonia in a complex reaction istration of NAG, since NAG undergoes cytosolic inacti-
that uses C02, NADH, N5 ,N10-methylenetetrahydrofolate vation by deacylation and is not readily permeable across
(a source of a single carbon unit; Chapter 27) and pyridoxal the inner mitochondrial membrane. An analogue of NAG,
345 17.3
Metabolism
N-carbamoylglutamate, activates CPSI, does not share the mediate in the urea cycle pathway and is also obtained
undesirable properties of NAG, and has been effective in from dietary proteins. A number of key metabolites such
management of this deficiency. as nitric oxide, phosphocreatine, spermine and ornithine
The most common cause of hyperammonemia in adults are derived from arginine. During normal growth and de-
is disease of the liver (e.g., due to ethanol abuse, infection, velopment, under certain pathological conditions (e.g., en-
or cancer). The ability to detoxify ammonia is decreased dothelial dysfunction) and if the endogenous production
in proportion to the severity of the damage. In advanced of arginine is insufficient, a dietary supplement of arginine
disease (e.g., cirrhosis), hyperammonemia is augmented may be required. Thus, arginine is considered a semi essen-
by shunting of portal blood that carries ammonia from the tial amino acid.
intestinal tract and other splanchnic organs to the systemic
blood circulation (bypassing the liver) and leads to portal-
systemic encephalopathy. In addition to dietary protein Metabolism and Synthesis of Nitric
restriction, colonic growth of bacteria must be suppressed Oxide
by antibiotics (e.g., neomycin) and administration of lac-
Nitric oxide (NO) is a reactive diatomic gaseous
tulose (Chapter 9), a nonassimilable disaccharide. Enteric
molecule with an unpaired electron (a free radical). It is
bacteria catabolize lactulose to organic acids that convert
lipophilic and can diffuse rapidly across biological
NH3 to NHt, thereby decreasing absorption of NH3 into
membranes. NO mediates a variety of physiological
the portal circulation. Catabolism of lactulose also leads to
functions such as endothelial derived relaxation of
formation of osmotically active particles that draw water
vascular smooth muscle, inhibition of platelet
into the colon, produce loose, acid stools, and permit loss
aggregation, neurotransmission, and cytotoxicity. The
of ammonia as ammonium ions.
pathophysiology of NO is a double- edged sword.
Insufficient production of NO has been im- plicated in
the development of hypertension, impotence,
17.3 Metabolism of Some Individual Amino Acids
susceptibility to infection, and atherogenesis. Excessive
NO production is linked to septic shock, inflamma-
Mammalian tissues synthesize the nonessential amino
tory diseases, transplant rejection, stroke, and carcino-
acids from carbon skeletons derived from lipid and car-
genesis.
bohydrate sources or from transformations that involve
NO is synthesized from one of the terminal nitrogen
essential amino acids. The nitrogen is obtained from
atoms or the guanidino group of arginine with the con-
NH! or from that of other amino acids. Nonessential
comitant production of citrulline. Molecular oxygen and
amino acids (and their precursors) are glutamic acid
NADPH are cosubstrates and the reaction is catalyzed
(o-ketoglutaric acid), aspartic acid (oxaloacetic acid),
by nitric oxide synthase (NOS). NOS consists of sev-
serine (3-phosphoglyceric acid), glycine (serine), tyro-
eral isoforms and is a complex enzyme containing bound
sine (phenylalanine), praline (glutamic acid), alanine
FMN, FAD, tetrahydrobiopterin, heme complex, and non-
(pyruvic acid), cysteine (methionine and serine), arginine
heme iron. A calmodulin binding site is also present. NO
(glutamate-y-semialdehyde), glutamine (glutamic acid),
formation from arginine is a two-step process requiring
and asparagine (aspartic acid).
five-electron oxidations. The first step is the formation of
Amino acids may be classified as ketogenic, glucogenic,
NG-hydroxylarginine (NG denotes guanidinium nitrogen
or glucogenic and ketogenic, depending on whether feed-
atom):
ing of a single amino acid to starved animals or animals
with experimentally induced diabetes increases plasma or Arginine+ 02 + NADPH + H+ -+
urine levels of glucose or ketone bodies (Chapter 18).
HO-NG-Arg + NADP+ + H20
Leucine and lysine are ketogenic; isoleucine, phenylala-
nine, tyrosine, and tryptophan are glucogenic and keto- This step is a mixed-function oxidation reaction simi-
genie; and the remaining amino acids are glucogenic. lar to the one catalyzed by cytochrome P-450 reduc-
Points of entry of amino acids into the gluconeogenic path- tase and there is considerable homology between NOS
way are discussed in Chapter 15. and cytochrome P-450 reductase. In the second step, fur-
ther oxidation of NG-hydroxyl arginine yields NO and
citrulline:
1
Arginine HO-NG-Arg + 02 + -(NADPH + H+)-+
2
Arginine participates in a number of metabolic pathways 1
citrulline + NO+ H20 + NADP+
depending on the cell type. It is synthesized as an inter- 2
346 17.3
Metabolism
The overall reaction is: interferon-y ), or bacterial lipopolysaccharides can

O H NH2 induce and cause expression of NOS in many cell
II I I
-Q-C-CH-(CH) -N-C=NH + 20
I + NADPH types. Glu- cocorticoids inhibit the induction of iNOS.
+W
23 In stimulated
macrophages and neurophils, NO and superoxide
radi-
NH3+
cal (02) react to generate peroxynitrite, a powerful
Arginine
ox- idant, and hydroxyl radicals. These reactive
Nitric Oxide FMN, FAD, intermedi-
Synthase Tetrahydrobiopterin
Fe2+, Heme
j complex
ates are involved in the killing of phagocytized
bacteria (Chapter 14). Excessive production of NO due
to endotox- inemia produces hypotension and vascular
hyporeactivity to vasoconstrictor agents, and leads to
septic shock. NOS
O H O
II I II inhibitors have potential therapeutic application in
-o- C - CI H - (CH 2 ) 3 - N - C - NH 2 + the treatment of hypotensive crisis.
NO
NH3+
Citrulline
Signal Transduction of NO
The NOS activity is inhibited by NG-substituted
analogues of arginine, such as NG-nitroarginine NO is lipophilic and diffuses readily across cell
and NG- monomethyl-L-arginine. membranes. It interacts with molecules in the target
cells pro- ducing various biological effects. One
mechanism of action of NO is stimulation of
Isoforms (Also Known as Isozymes)
guanylate cyclase, which catalyzes the formation of
of Nitric Oxide Synthase
cyclic guanosine monophosphate (cGMP) from
There are three major isoforms of Nitric GTP, resulting in increased intracellular cGMP levels
Oxide (Figure 17-9). NO activates guanylate cyclase by
Synthase (NOS) ranging in molecular size from 130 binding to heme iron. The elevation of cGMP levels
to
may activate cGMP-dependent protein kinases.
160 kDa. Amino acid similarity between any two
isoforms is about 50-60%. Isoforms of NOS exhibit
0
differences in tissue distribution, transcriptional
regulation, and activation by intracellular Ca2+. Two
of the three isoforms of
HN:.r\_ 0 0 0
NOS are constitutive enzymes (cNOS) and the third I / H II II II
iso-
H' N J,,,N �c'-o- P- o-I P- o-I P-o-
forin is an inducible enzyme (iNOS). The cNOS O 5 I
H H H 3· H 0- 0- 0-
isoforms are found in the vascular endothelium
0H OH
(eNOS), neuronal cells (nNOS), and many other
�:;::r��uTse
'" 1
cells, and are regulated by Ca2+ and calmodulin. In Guanosine Triphosphate (GTP)
the vascular endothelium, agonists such as

acetylcholine and bradykinin activate eNOS by Guanylate Cyclas •• ----0---- ,o (
enhancing intracellular Ca2+ concentrations via the
O Derived from
production of inositol 1,4,5-trisphosphate, which a
activates the phosphoinositide second-messenger HN�N;> Ligand-cell interaction
system (Chapter 30 ). The NO produced in the vascular ,l,,,�.Jl H2

endothelium maintains basal vascular tone by H,N N �·o,
H H H H � .-< 0 Smooth muscle
vasodilation which is mediated by vascular smooth 3' P� ...._ relaxation
muscle cells. Organic nitrates used in the management of

OH 0/ 'o- O
Cyclic GMP HN�N�CH,
ischemic heart disease act by denitration with the
-----0-
y
I
Cyclic GMP CH,C�N�
subsequent formation of NO. Sodium nitroprusside, an phosphodiesterase CH CH CH
antihypertensive drug, is an NO
donor. Thus, organic nitrates and sodium nitroprusside are prodrugs, and the which prodr
exact mechanism by these ugs
347 17.3
Metabolism
yield NO is not yet understood. Inhaled NO can produce 2 2 3
pulmonary vasodilation. This property of NO has been 0 o,s,Nl

used in the management of hypoxic respiratory failure �N,
H��N;> H � CH3
associated with primary pulmonary hypertension in H a N �NJlx0;j5 c'-o- PI - Sildenafil
neonates. NO produced by cNOS in neuronal tissue o-
functions as a neurotransmitter. H HH H o-
The inducible class of NOS (iNOS) is found in OH OH
5'·GMP
FIGURE 17-9
NO-mediated synthesis of cGMP from GTP in the corpus cavernosum that
macrophages and neutrophils and is Ca2+ -independent. leads to smooth muscle relaxation. Sildenafil potentiates the effects of NO
Bacterial endotoxins, cytokines (e.g., interleukin- by inhibiting cGMP phosphodiesterase.
I,
348
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 Protein and Amino Acid 347
Metabolism
U
Proteins Punrnes
Glutathione \_
.> f
3-Phosphoglycerate /
Glucose
rea------ '\_ r ------Pyruvat�
·�lntnx
"One-carbon... + NH;�;lyc\
' Serine� Ethanolamine- Cho::: cycle-co,
Bile acid conjugates / I \ <, l /

Hippurate and Glyoxalate- Glycolate
Phospholipids other acyl conjugates !
Porphyrins Creatine Oxalate
l
Phosphocreatin
e
l
Creatinine
FIGURE 17-10
Overview of glycine and serine metabolism.
These kinases phosphorylate specific proteins that The antiaggregability of platelets and the neurotoxicity
may be involved in removal or sequestration of Ca2+ of
or other ions, resulting in physiological stimuli. The NO have been attributed to inhibition of glycolysis by
physiological actions of cGMP are terminated by its NO.
conversion to
5' -GMP by cGMP-phosphodiesterase. Inhibitors of Glycine
cGMP
Glycine particrpates in a number of synthetic
phosphodiesterase promote the actions of
NO. pathways and is oxidized to provide energy (Figure
Sildena.fil is a selective inhibitor of a specific 17-10). The interconversion of glycine and serine
cGMP phosphodiesterase (type 5) present in the by serine hydroxymethyltransferase is shown below:
corpus cavernosum. This compound (structure shown pyridoxal phosphate
in Figure 17-9) is used orally in the therapy of some

types of erectile dys-
function. NO is the principal transmitter involved in NHj-CH2-COO- + N5,N -methylene-FH4 + H20 +====::::±
the
10
glycine
relaxation of penile smooth muscle. During central or
re- flex sexual arousal, NO production is enhanced serine
leading
to increased production of cGMP. Smooth muscle Signal transduction of NO by cGMP-
relaxation permits the corpus cavemosum to fill independent mechanisms include ADP-ribosylation of
with blood. Since the therapeutic effect of sildenafil glyceraldehyde-
potentiates the action of cGMP, the drug is ineffective 3-phosphate dehydrogenase (GADPH), an enzyme of
in the absence of sexual arousal. The relaxation of the glycolytic pathway (Chapter 13), and interactions
cavernosal smooth mus- with many heme-containing and nonheme iron-sulfur
cle caused by cGMP involves inhibition of Ca2+ containing proteins. NO activates ADP-
uptake. ribosyltransferase which catalyzes the transfer of
Prostaglandin E, (alprostadil) inhibits the uptake of ADP-ribose from NAD+ to GADPH. This results in
Ca2+ the inactivation of GADPH caus- ing inhibition of
smooth muscle by a separate mechanism and causes
glycolysis and decreased ATP production.
erec- tions in the absence of sexual arousal. Blood flow
through corpus cavernosum may also be increased by o-
adrenergic blocking agents (e.g., phentolamine
mesylate). Coadmin- istration of NO donor drugs with
the NO potentiation drug sildenafil may have severe
consequences on the cardiovascular system.
348
Metabolism
10
The one-carbon carrier N5 ,N -
methylenetetrahydrofolate is derived from reactions of
the one-carbon pool (Chap- ter 27). [The term
one-carbon pool refers to all single-carbon-
containing metabolites (e.g., -CH3, -CHO, NH=C-,
etc.) that can be utilized in biosynthetic reactions
such as formation of purine and pyrimidine.] These
reactions include oxidation of glycine by glycine
cleavage
enzyme complex (glycine
synthase):
pyridoxal phosphate
NHf-CH2-Coo- + FH4 + NAD+
+=======:±
glycine
NHt + C02 + NADH + N5,N10-methylene-FH4

This reaction favors glycine degradation, but the
formation of glycine may also occur. The enzyme
complex is mitochondrial and contains a pyridoxal
phosphate-dependent glycine decarboxylase, a
lipoic acid-containing protein that is a carrier
of an aminomethyl moiety, a tetrahydrofolate-
requiring enzyme, and lipoamide
dehydrogenase. The reactions of glycine cleavage
re- semble those of oxidative decarboxylation of
pyruvate (Chapter 13).
348
Metabolism
Glycine is also oxidized by n-amino acid oxidase, Creatine and Related Compounds
an
Phosphocreatine serves as a high-energy
FAD
phosphate donor for ATP formation (e.g., in muscle
protein:
contraction; see Chapter 21 ). Synthesis of creatine
NHj -CHrCOO- + 02 + H20 (methyl guanidinoacetate) requires transamidination,
� i.e., transfer of a guani- dine group from arginine to
glycine glycine, to form guanidinoacetate (glycocyamine) by
CHO-coo- + NHt + mitochondrial arginine-glycine amidinotransferase
H202 NH2
glyoxalate
Glyoxalate can be transaminated to glycine, reduced

to glycolate, converted to a-hydroxy-,B-ketoadipate by
reaction with a-ketoglutarate, or oxidized to oxalate
and ex- I NH2
creted in urine. The first three reactions require C=NH2+ I NH3 +
I C=NH2+ I
pyridoxal phosphate, NADH, and thiamine NH I (CH2b
pyrophosphate, respec- tively. In humans, ascorbic acid I NH + I
(CH2h I CHNH3
(vitamin C) is a precursor of urinary oxalate (Chapter I CH2 I
CHNH3 + I coo-
38). Since calcium oxalate is poorly soluble in water, coo-
I
it can cause nephrolithiasis and nephrocalcinosis due coo-
to hyperoxaluria.
Arginine Glycine Guanidinoacetate Ornithine

Disorders of Glycine Catabolism (glycocyamine)
Nonketotic hyperglycinemia is an inborn error due to
a
defect in the glycine cleavage enzyme complex in In the next step guanidinoacetate is methylated by
which glycine accumulates in body fluids and S- adenosylmethionine by cytosolic S-
especially in cerebrospinal fluid. It is characterized adenosylmethionine guanidinoacetate-N-
by mental retardation methyltransferase to form creatine.
and seizures. Glycine is an inhibitory
CH3 NH2 NH2
neurotransmitter
in the central nervous system, including the spinal cord. I S·Adenosyl
I I
Strychnine, which produces convulsions by competitive S + -Adenosyl C=NH2 + C=NH2+ I
I I I (CH2h
inhibition of glycine binding to its receptors, gives mod- (CH2h + NH N-CH3 + I +
I CHNH3 + H+
est results in treatment but not very effective. Sodium I I
CHNH3 + CH2 CH2 I
benzoate administration reduces plasma glycine levels I I I coo-
but does not appreciably alter the course of the coo· coo· coo·
disease. Ex-
change transfusion may be useful. Ketotic S-Adenosyl- Guanidino- Creatine S-Adenosyl-
methionine acetate homocysteine
hyperglycinemia also occurs in propionic acidemia
but the mechanism
has not been established.
Primary hyperoxaluria type I is due to a deficiency
O OH
of cytosolic a-ketoglutarate-glyoxylate carboligase, II I
which catalyzes the following reaction: C02+ -ooc-CH2-CH2-C-CH-
COO-
CX·Hydroxy-{J-ketoadipate
The glyoxylate that accumulates is converted

Glyoxalate cx-Ketoglutarate to oxalate.
Carboligase Thiamine pyrophosphate

348
Metabolism
These reactions occur in liver, kidney, and pancreas,
from which creatine is transported to organs such as
muscle and brain. Creatine synthesis is subject to
negative modu- lation of amidinotransferase by
creatine. Phosphocreatine production is catalyzed by
creatine kinase:
NH2 NH-PO/-
I I
C=NH2 + C=NH2 +
I I
N-CHa + ATP4- N-CH3 + ADP3- + H+
I I
CH2 CH2
I I
coo- coo-
Creatine Phosphocreatine
This kinase is a dimer of M and B (M = muscle, B

= brain) subunits produced by different structural
genes. Three isozymes are possible: BB (CK-1), MB
(CK-2), and MM (CK-3). Another isozyme differs
immunologically and electrophoretically and is
located in the inter- membrane space of mitochondria.
Tissues rich in CK-1
350
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 349
are brain, prostate, gut, lung, bladder, uterus, regenerates ATP from ADP, thereby maintaining a
placenta, and thyroid; those rich in CK-3 are high level of ATP required during intense exercise. A
skeletal and cardiac muscle. Cardiac muscle contains large pool of phosphocreatine resides in the skeletal
significant amounts of CK-2 (25-46% of total CK muscle. It has been theorized that in order to maximize
activity, as opposed to less than 5% in skeletal phosphocreatine stores in the skeletal muscle to
muscle), so that in myocardial infarction the rise in replenish ATP during rapid muscle contractions, an
serum total CK activity is accompa- nied by a parallel exogenous source of creatine may be beneficial.
rise in that of CK-2 (Chapter 8). Double-blind placebo-controlled studies of oral
Phosphocreatine undergoes a slow and sup-
nonenzymatic cyclization to creatinine. plementation of creatine in human subjects have
shown increased performance during short duration,
NH- strenuous, high-intensity exercise. Such activities
PO/- require that ATP be replenished rapidly from
I phosphocreatine stores during anaerobic metabolism.
C=NH2
+ These studies usually consisted of ingestion of 20 g of
I creatine per day for 5 days followed by a maintenance
N-CH3 +H•-
dose of 5-10 g/day. Studies on creatine as an
1 ergogenic aid have not been uniformly positive;
CH2
some have shown no beneficial effect and still others
I have
coo-
Phosphocreatine Creatinine been equivocal and indicated that creatine
supplementation did not enhance athletic activities.
The safety issues
Creatinine has no useful function and is eliminated of long-term creatine supplementation on kidney,
by renal glomerular filtration and to a small extent by liver, nerve, muscle, and other tissues are not known.
renal tubular secretion. Creatinine clearance
approximately par- allels the glomerularfiltration rate Serine
(GFR) and is used as a kidney function test. It is
calculated as follows: Synthesis of serine from 3-phosphoglycerate, an
intermediate of glycolysis (Chapter 13), requires
. . urine creatinine (mg/L) oxidation of 3-phosphoglycerate to 3-
Creatinine clearance = --------
- phosphohydroxypyruvate, transamination of 3-
plasma creatinine (mg/L)
phosphohydroxypyruvate by glutamate, and
x urine volume per unit
hydrolysis of 3-phosphoserine to serine (Figure 17-
time
11). This cytosolic pathway is regulated by
Creatinine concentrations are measured from a inhibition of phosphoserine phosphatase by serine.
pre- cisely timed urine specimen (e.g., 4-hour, 24- Serine is converted to pyruvate by cytosolic serine
hour) and a plasma specimen drawn during the dehydratase. More importantly, it is converted in
urine collection period. Excretion of creatinine mitochondria to
depends on skeletal muscle mass and varies with 2-phosphoglycerate by way of hydroxypyruvate
age and sex. However, day-to- day variation in a and n-glycerate; and the enzymes involved are a
healthy individual is not significant. Creatinuria, the transaminase, a dehydrogenase, and a kinase.
excessive excretion of creatine in urine, may occur Serine is in- terconvertible with glycine (Figure
during growth, fever, starvation, diabetes melli- tus, 17-10) and is involved in phospholipid (Chapter
extensive tissue destruction, muscular dystrophy, and 19) and in cysteine
hyperthyroidism. synthesis
.
Use of Creatine as a Dietary Supplement glycine, arginine, and methionine, creatine is
synthesized in the liver, pancreas, and kidney. Creatine
The creatine pool in the human body comes from transported in blood crosses muscle and nerve cell
both endogenous synthesis and the diet which provides membranes by means of a specific creatine
1-2 g. Red meat provides large amounts of dietary transporter system against a concentration gradient of
creatine and vegetables a limited amount. Using
350
200: 1. Intracellularly, creatine is converted to Proline
phosphocre-
atine by ATP, a reaction catalyzed by creatine kinase. Proline arises from and gives rise to glutamate.
Phos- phocreatine, with its high phosphoryl transfer Synthesis is by reduction of glutamate to glutamate-y-
potential, semialdehyde by way of an enzyme-bound y-
glutamyl phosphate. The y-semialdehyde
spontaneously cyclizes to l::/-pyrroline-
5-carboxylate, which is then reduced by NAD(P)H to
proline (Figure 17-12). Proline is converted to zx' -
pyrroline-5- carboxylate by proline oxidase, which is
tightly bound to the inner mitochondrial membrane in
liver, kidney, heart,
and brain. t::.' -Pyrroline-5-carboxylate is in
equilibrium
with glutamate-y-sernialdehyde, which can be
transaminated to ornithine or reduced to glutamate
(Figure 17-12).
350
coo- coo-
Phosphoglycerat
Glycolysis--
l
HC-OH e l
- I de�rogenase r=o
H2C-o-® H,C-O-®
3- 3-Phosphohydroxypyruvate
Phosphoglycerate
Glutamate
osphoserin
e
nsaminase
a-Ketoglutarate
�
coo- coo-
l Phosphoserin
+H NyH e
3
phosphatase H2N?H
CH;- ? '\HO
fl
H,c-o-®
OH 3-
Phosphoserine
FIGURE 17- L·Serine
11
Synthesis of serine from 3-phosphoglycerate. P =PO�-; P; = HPO�-.
Decarboxylation of omithine to putrescine by omithine decarboxylase. Omithine aminotransferase deficiency is

decarboxylase serves as a source of the polyamines associated with gyrate atrophy of the choroid and retina
spermidine and spermine. Ornithinemia results from (Chapter 38). Praline and hydroxyproline (produced by
deficiency of omithine aminotransferase or ornithine posttranslational modification) are major constituents
coo-
l
(1H,�
ATP
CHNH;
ADP+ fl ,----� I
coo-
NADPH+H CD L-Glutamate
+
CHO_),
(6H,� @ NADH +
H+
I
CHNH+ NAO
+
I '
coo- H,C--CH,
Glutamate- Y - I . I
semialdehyde HC"'-'::N/CHCOOH
!,-- Glutamate t:,. -Pyrroline-

®!'-- a -Ketoglutarate 5-carboxylate
CH,NH:
I NAD(P)H
H+ +
(CH,� co,
CHNH;
I
tis> CH,-
NH; NAD(Pt
@ Flavoprotein
0,
co-r I
0rnithine (CH,),
i Via I H,C--CH,
CH,NH: I I
Put reseine
H,C......_N/CHCOOH
i reactions
of H
i urea cycle Proline
Arginine
FIGURE 17-12
Metabolism of proline (I) 6'-pyrroline-5-carboxylate (P5C) synthase; (2) ornithine aminotransferase; (3) PSC
reductase; (4) proline oxidase; (5) PSC dehydrogenase; (6) ornithine decarboxylase.
352
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11Protein and Amino Acid 351
Metabolism
of collagen (Chapter 25). Hydroxyproline released by N5-methyltetrahydrofolate and is unavailable as a carrier

collagen turnover undergoes degradation similar to for the formimino group of Figlu.
that of proline. Hydroxyproline cleavage initiated by
Homocysteine methyttransferase
hydroxyproline oxidase eventually yields glyoxylate and 5 (methylcobal amin )
pyruvate. In hyperprolinemia type I, proline
N -Methyl-FH, ---' - " '-'-----+
FH,
oxidase is deficient, and in type II, 6' -pyrroline-5- 7
Homocysteine Methionine
carboxylate dehydrogenase is deficient.

Hydroxyprolinemia results from hydroxyproline Similarly, a deficiency of glutamate formiminotransferase
oxidase deficiency. All are clinically harmless autosomal leads to accumulation of Figlu and high levels of serum
recessive traits. folate.
Histidinemia results from deficiency of
Histidine histidine ammonia-lyase. With a normal diet, histidine
(and the products imidazole-pyruvate, imidazole-
Histidine is not essential for adults except lactate, imidazole- acetate) accumulates in plasma,
in persons with uremia. It is essential for growth cerebrospinal fluid, and urine. This rare autosomal
in children. Histidine is synthesized from 5- recessive disease may be benign or may manifest with
phosphoribosyl- mental retardation and speech defects.
1-pyrophosphate and ATP, forming N' -1 '-phosphoribosyl- Histidine and ,B-alanine yield the dipeptide carnosine
ATP, catalyzed by the allosteric enzyme ATP phosphori- (present in muscle), and histidine and y-aminobutyrate
bosyltransferase. This reaction is analogous to the initial yield homocarnosine (found in brain). Methylhistidyl resi-
reaction of purine nucleotide biosynthesis (Chapter 27). dues are found in some proteins (e.g., actin; Chapter 21)
Histamine breakdown produces a one-carbon unit (N5-
as a result of posttranslational modification. Histamine is
formiminotetrahydrofolate) and glutamate (Figure 17-13)
decarboxylated histidine.
by a nonoxidative deamination to urocanate, cleavage of
the imidazole ring to N-formiminoglutamate (Figlu), and Histidine decarboxytase
transfer of the formimino group (-CH=NH) to tetrahy- (pyridoxal
phosphate)
drofolate (Chapter 27). I I I CH2-CH2-NH3 +
Folate deficiency leads to accumulation of Figlu, which
is excreted in urine. The excretion is very pronounced '\co, N�NH
after a loading dose of histidine, a test used to detect fo-
late deficiency. More sensitive radioisotopic assays use fo- Histidine Histamine
late binders to the vitamins. High urinary levels of Figlu

Histamine occurs in blood basophils, tissue mast cells,
may coexist with elevated levels of serum folate. Thus, in
and certain cells of the gastric mucosa and other parts of
vitamin B12 (cobalamin) deficiency, since cobalamin
the body (e.g., anterior and posterior lobes of the pituitary,
participates in the following reaction, FH4 is trapped as some areas of the brain). Histamine is a neurotransmitter in
certain nerves C'histaminergic'') in the brain. In mast cells
found in loose connective tissue and capsules, especially
NH+
I ' Histidine-
around blood vessels, and in basophils, histamine is stored
I I CH,-CH-coo- ammonia·
ase I I CH==CHCOO- in granules bound by ionic interactions to a heparin-protein
N�N (histidase) N�NH complex and is released (by degranulation, vacuoliza-
H NH,
L·Histidin
e
Urocanate tion, and depletion) in immediate hypersensitivity reac-
HP� tions, trauma, and nonspecific injuries (infection, burns).
l Urocanat
O H e
II I hydratase O
tmidazolone- Degranulation is affected by oxygen, temperature, and
-o-c=====1-CH;-
CH,COO-
propionase
"\
I
0... I c H;--CH,CO -
metabolic inhibitors. Release of histamine from gastric
H,0 N�N
HN:::a:, ,,-NH H mucosa! cells is mediated by acetylcholine (released by
H
r 4-lmidazolone-5-
propionate parasympathetic nerve stimulation) and gastrin and stim-
N·Formiminoglutamate
(Figlu) ulates secretion of hydrochloric acid (Chapter 12). His-
Glut;,mate tamine causes contraction of smooth muscle in various
r:FH,
forrnirninc- organs (gut, bronchi) by binding to H1 receptors. The
transferase N'·formino-
FH. conventional antihistaminic drugs (e.g., diphenhydramine
0
- 0-C-?H-CH,CH,COO-
II
NH:
352
L -Glutamate Metabolism
and pyrilamine) are H 1-receptor antagonists and are use-
FIGURE 17-13 ful in the management of various allergic manifestations.
Catabolism of histidine. However, in acute anaphylaxis, bronchiolar constriction is
rapidly relieved by epinephrine (a physiological antagonist
352
Metabolism
of histamine). Its effect on secretion of hydrochloric acid Leucme. isoleuone. and

is mediated by H2 receptors. H2-receptor antagonists are val,ne
Glutamate
cimetidine and ranitidine (Chapter 12), which are useful Transarmnauon
in treatment of gastric ulcers. Histamine is rapidly inacti- a-
vated by methylation from S-adenosylmethionine of one Ketoglutarate
of the nitrogen atoms of the imidazole ring (catalyzed by Corresponding a ·keto acids
N-methyltransferase) or of the terminal amine group (cat- CoASH
alyzed by methyltransferase). Ring-methylated histamine Oxtdatlve decart>oxyla ion (TPP. hpoam,de.
is deaminated by monoamine oxidase to methyl imidazole FAD) CO,. NADH + H+
acetic acid, which is readily excreted. Inactivation also re- Corresponding acyl-CoA
sults from deamination of histamine by diamine oxidase. 1h1oes1ers
The imidazole acetic acid formed is then excreted as 1-
k-
ribosylimidazole-4-acetic acid. This reaction is the only ;��hydrogenat1on
known reaction in which ribose is used for conjugation. FFADH,
Corresponding a.p
-unsaturated
/acyl-CoA th1oesters
F,-ro-m-e_ucl.. ,. n� j From 1soleucinej

•
'" "
:
Branched-Chain Amino Acids ll·Hydroxy-r. -
methyl
Prop1onyl-CoA
and
Succmyl·CoA
gtutary -CoA acetyl·CoA
Leucine, isoleucine, and valine are essential amino acids
but can be derived from their respective o-keto acids. A Acetoaceta e
single enzyme may catalyze transamination of all three. and acetyl-CoA
The o-keto acids, by oxidative decarboxylation, yield FIGURE 17-14

the acyl-CoA thioesters, which, by a,/3-dehydrogenation, which inhibits this step in /3-oxidation, also inhibits
yield the corresponding a,/3-unsaturated acyl-CoA it in the catabolism of branched-chain amino acids.
thioesters. The catabolism of these thioesters then di-
verges. Catabolism of leucine yields acetoacetate and
acetyl-CoA via /3-hydroxy-/3-methylglutaryl-coenzyme
A (HMG-CoA)-also an intermediate in the biosynthe-
sis of cholesterol and other isoprenoids (Chapter 19).
Catabolism of isoleucine yields propionyl-CoA (a gluco-
genie precursor) and acetyl-CoA. Catabolism of valine
yields succinyl-CoA (Figure 17-14 ). Thus, leucine is
ketogenic and isoleucine and valine are ketogenic and
glucogenic.
Oxidative decarboxylation of the o-keto acids is cat-
alyzed by a branched-chain keto acid dehydrogenase
(BCKADH) complex analogous to that of the pyru-
vate dehydrogenase and o-ketoglurarate dehydrogenases
complexes. BCKADH is widely distributed in mam-
malian tissue mitochondria (especially in liver and kid-
ney). It requires Mg2+, thiamine pyrophosphate, CoA-SH,
lipoamide, FAD, and NAO+ and contains activities of a-
keto acid decarboxylase, dihydrolipoyl transacylase, and
dihydrolipoyl dehydrogenase. Like the pyruvate dehydro-
genase complex, BCKADH is regulated by product in-
hibition and by phosphorylation (which inactivates) and
dephosphorylation (which activates).
The a,/3-dehydrogenation is catalyzed by an FAD pro-
tein and is analogous to the dehydrogenation of straight-
chain acyl-CoA thioesters in /3-oxidation of fatty acids
(Chapter 18). Methylenecyclopropylacetyl-CoA derived
from the plant toxin hypoglycin (Chapters 15 and 18),
352
Overview of the catabolism of branched-chain amino acids. TPP = thiamin Metabolism
pyrophosphate.
Hypoglycin produces hypoglycemia and metabolic aci-

dosis, which frequently are fatal.
Branched-chain ketoaciduria (maple syrup urine
disease), an autosomal recessive disorder
characterized by ketoacidosis starts early in infancy and
is due to a defect in the oxidative decarboxylation
step of branched- chain amino acid metabolism. The
name derives from the characteristic odor (reminiscent
of maple syrup) of the urine of these patients. Five
different variants (classic, intermittent, intermediate,
thiamine-responsive, and dihydrolipoyl dehydrogenase
deficiency) are known, of which the first, which is due
to deficiency of branched- chain o-keto acid
decarboxylase, is the most severe. The incidence of
maple syrup urine disease in the U.S. population is I
in 250,000-400,000 live births. In Mennonite populations
the incidence is extremely high ( I in 760). Neonatal
screening programs consist of measuring leucine levels in
dried blood spots using a bacterial inhibition assay.
Neonatal screening programs usually include testing for a
number of other treatable metabolic disorders such as
hypothyroidism, phenylketonuria, galactosemia, and
others. If the screening test is positive for a given metabolic
disease, a confirmatory test is performed. For maple syrup
urine disease, the confirmation requires quantitation of
the serum levels of branched-chain amino acids and urine
levels of both the branched-chain amino acids and their
ketoacids. Long-term management includes dietary re-
striction of the branched-chain amino acids. Frequent
measurement of plasma concentrations of these amino
352
Metabolism
acids is necessary to monitor the degree of dietary restric-

tion and patient compliance.
+
Adenosine-S-(CH,�-CH-COOH
NH,
I
Adenosine-S-(CH,);-CH-COOH
r
I
Many aminoacidurias and their metabolites give rise CH,
S·Adenosylmethionine S·Adenosylhomocysteine
to abnormal odors, maple syrup urine disease is one ex- Specific methyltransferase
ample. Some of the others are phenylketonuria (musty
odor), tyrosinemia type I (boiled cabbage), glutaric Acceptor (examples) Acceptor - CH, (examples)
aciduria (sweaty feet), 3-methylcrotonylglycinuria ( cat's 1. Guanldinoacetic acid

2. Nicotinamide
1. Creatine
2. N·Methylnicotinamide
urine), and trimethylaminuria (fish). In patients with 3. Norepinephrine
4. Phosphatidylethanolamine
3. Epinephrine
4. Phosphatidylcholine
trimethylaminuria the compound responsible for the fish 5. N-Acetyl·serotonin (three cycles of methylation)
5. Melatonin
odor is trimethylamine which is a byproduct of protein FIGURE 17-15
catabolism by the large intestinal bacterial flora. Normally, Selected methyl transfer reactions involving S-adenosylmethionine.
trimethylamine is inactivated by hepatic flavin monooxy-
genases. Several different mutations in the gene for flavin
monooxygenases have been identified in trimethylamin- The methyl group is transferred to appropriate accep-
uric patients. An inhibitor of flavin monooxygenases is tors by specific methyltransferases with production of
indole-3-carbinol found in dark green vegetables (e.g., S-adenosylhomocysteine (Figure 17-15), which is hy-
broccoli). The amelioration of symptoms of bad odor in drolyzed to homocysteine and adenosine by adenosylho-
trimethylaminuria may be achieved by limiting intake of mocysteinase:
dark green vegetables and protein, and by administering
low doses of antibiotics to reduce intestinal bacterial flora. NH3 + NH3+
I _ H20 Adenosine I _
Adenosyl-S-(CH2)2-CH-COO HS-(CH2)2-CH-C00
S-Adenosylhomocysteine Homocysteine
Sulfur-Containing Amino
Acids Homocysteine can be recycled back to methionine
either by transfer of a methyl group from betaine catalyzed
Methionine and cysteine are the principal sources of or- by betaine-homocysteine methyltransferase, or from N5-
ganic sulfur in humans. Methionine is essential (unless
methyltetrahydrofolate (N5-methyl-FH4) catalyzed by N5-
adequate homocysteine and a source of methyl groups are
methyl-Fl-la-methyltransferase, which requires methyl
available), but cysteine is not, since it can be synthesized
cobalamin:
from methionine.
(1)
Methionine
Methionine is utilized primarily in protein synthe- Homocysteine Betaine
sis, providing sulfur for cysteine synthesis, and is the
NH3 +
body's principal methyl donor. In methylation reactions, I
S-adenosylmethionine (SAM) is the methyl group donor. H3C-S-(Ch,l,-CH-C00- + (CH3),N-CH,-coo-
Methionine Dimethylglycine
SAM is a sulfonium compound whose adenosyl moiety is
derived from ATP as follows:
(2)
NH3 + N5·Methyl-FH,-homocysteine
I methyltransferase
NH3 + HS-(CH 2) -CH-COO-
2
+ N5-Methyl-FH, (methylcobalamin)
I ATP,H,O PP,+P,
CH3-S-(CH2)2-CH-COO ,.,....,-,--�-----....
Methionine
adenosyltransferase
Methionine
Betaine (an acid) is obtained from oxidation of choline

(an alcohol) in two steps:
FAO FAOH2
(CH3h •N-CH2-CH20H :::,,, ,/ • (CH3h •N-CH2-CHO
Choline o>cidase
Choline Betaine aldehyde
NAO• lSetaine aldehyde

dehyctrogenaso
NAOH+H*
S-Adenosylmethionine (CH3l3N-CH,-Coo-
(active methionine) Betaine
354
Metabolism
Cysteine carboxylic acid group. Taurine is conjugated with bile

In the biosynthesis of cysteine, the sulfur comes from acids in the liver (Chapter 19) and is readily excreted by the
methionine by transsulfuration, and the carbon skeleton kidney. It is a major free amino acid of the central nervous
and the amino group are provided by serine (Figure 17-16). system (where it may be an excitatory neurotransmitter)
Cysteine regulates its own formation by functioning and the most abundant in the retina; it also occurs in other
as an allosteric inhibitor of cystathionine y-lyase. a- tissues (e.g., muscle, lung).
Ketobutyrate is metabolized to succinyl-CoA by way of Sulfate can be converted to the sulfate donor compound
propionyl-CoA and methylmalonyl-CoA. 3' -phosphoadenosine-5' -phosphosulfate (PAPS) in a two-
Cysteine is required for the biosynthesis of glu- step reaction (Figure 17-17). PAPS participates in the
tathione and of CoA-SH. A synthetic derivative, N- sulfate esterification of alcoholic and phenolic functional
acetylcysteine, is used to replenish hepatic levels of glu- groups (e.g., in synthesis of sulfolipids and glycosamino-
tathione and prevent hepatotoxicity due to overdosage glycans).
with acetaminophen. When high concentrations of ace-
toaminophen are present in the liver, the drug undergoes
Abnormalities Involving Sulfur-Containing
N-hydroxylation to form N-acetyl-benzoquinoneimine,
Amino Acids
which is highly reactive with sulfhydryl groups of pro-
teins and glutathione and causes hepatic necrosis. N- Deficiencies of methionine adenosyltransferase, cys-
Acetylcysteine is used as a mucolytic agent (e.g., in cystic tathionine ,'3-synthase, and cystathionine y-lyase have
fibrosis) because it cleaves disulfide linkages of mucopro- been described. The first leads to hypermethioninemia
teins. Cysteine and cystine are interconverted by NAD- but no other clinical abnormality. The second leads to hy-
dependent cystine reductase and nonenzymatically by an permethioninemia, hyperhomocysteinemia, and
appropriate redox agent (e.g., GSH). homocystinuria. The disorder is transmitted as an
The major end products of cysteine catabolism in hu- autosomal recessive trait. Its clinical manifestations
mans are inorganic sulfate, taurine, and pyruvate. Taurine may include skeletal abnormalities, mental retardation,
is a ,B-amino acid that has a sulfonic acid instead of a ectopia lentis (lens dislocation), malar flush, and
susceptibility to arte- rial and venous thromboembolism.
Some patients show reduction in plasma methionine and
L·Meth�
oni�� homocysteine concentrations and in urinary
P homocysteine excretion after large doses of pyridoxine.
thionine adenosyltransferase Homocystinuria can also result from a deficiency of
PP, + P.
cobalamin (vitamin B12) or folate metabolism. The
S·Adenosylmethionin
e third, an autosomal recessive trait, leads to
cystathioninuria and no other characteristic clinical
� (- CH,) in methyltransferase abnormality.
reaction Hereditary sulfite oxidase deficiency can occur alone
S·Adenosy�
hom�steine or with xanthine oxidase deficiency. Both enzymes con-
enosylhomocysteinas tain molybdenum (Chapter 27). Patients with sulfite ox-
e idase deficiency exhibit mental retardation, major motor
Adenosine seizures, cerebral atrophy, and lens dislocation. Dietary
HS-CH,-CH,-rH- deficiency of molybdenum (Chapter 37) can cause defi-
COO-
cient activity of sulfite and xanthine oxidases.
NH,+ • Cystinuria is a disorder of renal and gastrointestinal
L -Homo�steine • 7H,+ • tract amino acid transport that also affects lysine, or-
HOH,C-t:H-COO- L·Serine nithine, and arginine. The four amino acids share a com-
stathionine � -synthase (pyridoxal
phosphate) H,O
NH+
- • I. , •
OO - H,-S-CH-c�-cH-COO-
' . .. I
C CH-C
NH+
mon transport mechanism (discussed above). Clinically, it
presents as urinary stone disease because of the insolubil-
L -Cysta�
hio�� '
ity of cystine. In cystinosis, cystine crystals are deposited
stathionine Y -lyase (pyridoxal
phosphate) NH/ in tissues because of a transport defect in ATP-dependent
(cystathionase) cystine efflux from lysosomes (discussed above).
NH,+ 0
• I. •
- OOC-CH-CH,-SH II
CH,-CH,-c-coo- - - - succinyl-CoA
+
354
L·Cysteine a·Ketobutyrate
Metabolism
Homocysteine
FIGURE 17-16
Biosynthesis of cysteine. The sulfur is derived from methionine, and the Homocysteine is an amino acid not found in pro-
carbon skeleton and amino group are derived from serine. teins. Its metabolism involves two pathways; one is
354
Metabolism
o-w-o ,��rase -o-M-o-[-o-p

- � -
N
<"
Sulfate
x5 N
II I '
0 ATP PP, O o-
HO OH
Adenosine 5 · -phosphosulfate
Adenylylsulfate
ATP
kinase Mg'+
ADP
-� � d<x5
o-s-O-P-0-CH,
II
0 0
I_
O
0
OH
-O=P-o-
I
6-
PAPS
FIGURE 17-17
Formation of 3' -phosphoadenosine-5' -phosphosulfate (PAPS).
the methylation of homocysteine to methionine remethylation or transsulfuration pathways. Individuals

using N5-methyltetrahydrofolate (N5-methyl-FH4) with a homozygous defect in cystathionine
catalyzed by a vitamin B 12-dependent enzyme. The
second is the transsulfuration pathway where
homocysteine condenses with serine to form
cystathionine; this is catalyzed by cystathionine ,B-
synthase (CBS) which is a pyridoxal-
5'-phosphate enzyme. End products of the
transsulfuration pathway are cysteine, taurine,
and sulfate (Figure 17-18). The methyl donor N5-
methyl-FH4 is synthesized from N5,N10-methylene-
FH4 and the reaction is catalyzed by N5 ,N10-
methylenetetrahydrofolate reductase (MTHFR).
MTHFR is a FAD-dependent enzyme. Thus, the
metabolism of homocysteine involves four water sol-
uble vitamins, folate, vitamin B12, pyridoxine, and
ri- boflavin. Any deficiencies or impairment in the
conversion of the four vitamins to their active
coenzyme forms will affect homocysteine levels.
Severe cases of hyperhomocysteinemia occur due to
deficiencies of enzymes in the homocysteine
354
Metabolism
,B-synthase have severe hyperhomocysteinemia
(plasma concentrations >50 µ.MIL) and their clinical
manifestations are premature atherosclerosis,
thromboembolic complications, skeletal abnormalities,
ectopia lentis and mental retardation.
In plasma, homocysteine is present as both free ( < 1
% ) and oxidized forms (>99%). The oxidized forms
include protein (primarily albumin)-bound
homocysteine mixed disulfide (80-90%),
homocysteine-cysteine mixed disulfide (5-10%), and
homocystine (5-10%). Several studies have shown the
relationship between homocysteine and altered
endothelial cell function leading to thrombosis. Thus,
hyperhomocysteinemia appears to be an independent
risk factor for occlusive vascular disease. Five to ten
percent of the general population have mild
hyperhomocysteinemia.
It has been shown that a thermolabile form of
MTHFR is a major cause of mildly elevated plasma
homocysteine levels, which have been associated
with coronary heart disease. The thermolabile
MTHFR gene has a
356
Metabolism
Folate (F)
2 NADPH+2H+
ydrofolate reductase (DR)

2 NADP+
Tetrahydrofolate (FH4)
Methyl
Serine S- acceptor
adenosylmethionine s
Serine
hydroxymethyl
transferase
(pyridoxal phosphate) Methylate
d acceptor
Glycine
Methylene FH4 Methylation

Methionine S-adenosylhomocysteine
Cycle
Methyl FH4
! MTHF
R
(FAD)
-------
Methyl
transferase
(methylcobalamin) Adenosyl-
homocysteinas
e
Homocysteine
µ Serine
ystathionine
Synthase yridoxal
phosphate)
Cystathionine
Transulfuration
Pathway
o-
+1j Cystathionine y-lyase
(pyridoxal phosphate)
Ketobutyrate
Cysteine
I \
I
Glutathione
\ Taurine---+ ---+ Sulfate
FIGURE 17-18
Homocysteine
metabolism.
mutation of C to T at nucleotide positron 677 which further studies are required to assess the utility of vitamin
causes an alanine-to-valine amino acid substitution in the supplementation.
protein. The mechanism by which homocysteine medi-
ates vascular pathology remains to be understood. The
targets for homocysteine damage are connective tissue,
Phenylalanine and
Tyrosine
endothelial cells, smooth muscle cells, coagulation fac-
tors, nitric oxide metabolism, plasma lipids and their Phenylalanine is an essential amino acid. Tyrosine is syn-
oxidized forms (Chapter 20). Vitamin supplementation thesized by hydroxylation of phenylalanine and there-
with B12, folate, and B6 has reduced total plasma homo- fore is not essential. However, if the hydroxylase sys-
cysteine levels. Vitamin supplementation may decrease tem is deficient or absent, the tyrosine requirement must
the morbidity and mortality from atherosclerotic vas- be met from the diet. These amino acids are involved in
cular disease due to hyperhomocysteinemia. However,
356
Metabolism
synthesis of a variety of important compounds, includ-
ing thyroxine, melanin, norepinephrine, and epinephrine
356
Metabolism
DIET
o-v- NH+ / � NH;
Pheny1a1anine
(essential)
�
�
.> HO-OCH,-bH-coo-
Ty�osine (essential
1� the event
of
inadequate
phenylalanine
supply)
::{n -: �Oxidation
/ (liver)
Thyroxine .
(thyroid gland)
Melanin
(skin pigment)
Neurotransmitters and [
Norepinephrine adrenal medullary
hormones Epine;hrine
FIGURE 17-19
Overview of the metabolism of phenylalanine and tyrosine.
(Figure 17-19). The conversion of phenylalanine to tyro- reductase is distributed widely in tissues (e.g., brain,
sine and its degradation to acetoacetate and fumarate are adrenal medulla).
shown in Figure 17-20. Human liver phenylalanine hydroxy lase is a multimeric
The phenylalanine hydroxylase reaction is complex, homopolymer whose catalytic activity is enhanced by
occuring principally in liver but also in kidney. The hy- phenylalanine and has a feed-forward metabolic effect.
droxylating system is present in hepatocyte cytosol and Phosphorylation of phenylalanine hydroxylase by cAMP-
contains phenylalanine hydroxylase, dihydropteridine re- dependant kinase leads to increased enzyme activity and
ductase, and tetrahydrobiopterin as coenzyme. The hy- dephosphorylation has an opposite effect. Thus, glucagon
droxylation is physiologically irreversible and consists of and insulin have opposing effects on the catalytic activity
a coupled oxidation of phenylalanine to tyrosine and of of phenylalanine hydroxylase.
tetrahydrobiopterin to a quinonoid dihydroderivative with Quinonoid dihydrobiopterin is an extremely unstable
molecular oxygen as the electron acceptor: compound that can rapidly rearrange (by tautomerization)
phenylalanine hydroxylase to 7,8-dihydrobiopterin (Figure 17-21) and be reduced to
Phenylalanine+ 02 + tetrahydrobiopterin----- the tetrahydro form by dihydrofolate reductase:
--
tyrosine + H20 + quinonoid-dihydrobiopterin dihydrofolate reductase
7,8-Dihydrobiopterin + NADPH + tt+------
The tetrahydrobiopterin is regenerated by reduction of the NADP+ + tetrahydrobiopterin
quinonoid dihydrobiopterin in the presence of NAD(P)H
by dihydropteridine reductase: This enzyme also catalyzes conversion of dihydrofolate
(FH2) to tetrahydrofolate (Fl-la), and folic acid contains a
Quinonoid dihydrobiopterin + NAD(P)H + pteridine ring system (see the discussion of one-carbon
dihydropteridine reductase
tt+ NAD(P)+ + tetrahydrobiopterin metabolism in Chapter 27). However, regeneration of
tetrahydrobiopterin by the dihydrofolate reductase reac-
NADH exhibits a lower Km and higher Vmax for tion, however, is too slow to support normal rates of pheny-
the reductase than NADPH. Thus, the pterin coen- lalanine hydroxylation.
zyme functions stoichiometrically (in the hydroxylase Tetrahydrobiopterin is synthesized starting from GTP
reaction) and catalytically (in the reductase reaction). and requires at least three enzymes. The first committed
Deficiency of dihydropteridine reductase causes a step is GTP-cyclohydrolase, which converts GTP to dihy-
substantial decrease in the rate of phenylalanine hy- droneopterin triphosphate. 6-Pyruvoyltetrahydrobiopterin
droxylation. Dihydropteridine reductase and tetrahydro-
synthase transforms dihydroneopterin triphosphate
biopterin are involved in hydroxylation of tyrosine and
into 6-pyruvoyltetrahydrobiopterin. The latter is reduced
of tryptophan to yield neurotransmitters and hormones
to tetrahydrobiopterin by NADPH-dependent sepi-
(dopamine, norepinephrine, epinephrine, and serotonin).
apterin reductase. Deficiency of GTP-cyclohydrolase and
Unlike phenylalanine hydroxylase, dihydropteridine
358
NH+
I • -0 , NH+
Metabolism
O
Q. Phenylalanine H,O
CH;-CH-coo- )> hydroxylase

Tetrahydro- Quinonoid·
< HO 'I
-
' CH,-JH-coo-
Phenylalanine biopterin dihydrobiopterin Tyrosine
t.
NAD(P} NAD(P)H+H+J
\ I
Dihydropteridine
a -Ketoglutarate
osine transaminase
Dihydrofolate reductase Glutamate
reductase NADP+ �
NADPH+H+
7,8-Dihydrobiopterin
p-Hydroxyphenylpyruvate
Hydroxylation, shift� H�roxyphenylpyruvic

of the side chain. acid oxidase; ascorbic
decarboxylation acid; Cu,.
co,
HO-q-OH
coo- CH,
l I CH,COO-
CH C=O
Homogentisic acid
II I
HC CH, O,
600-
Fumarate
600-
Acetoacetate
omogentisic acid
idase; ascorbic
id;GSH
�
r
H.0 Fumarylacetoacetase
-OOC /H coo-
\II l
HIT
C CH, CH Maleylacetoacetate HC CH\ CH,
1 \!
H II
\j
II
\oo-
isomerase
\I c! "coo-
0 0 II II
0 0
Fumarylacetoacetate Maleylacetoacetate
FIGURE 17-20
Conversion of phenylalanine to tyrosine and the oxidative pathway of tyrosine.
6-pyruvoyl tetrahydrobiopterin synthase leads to (types II and III). Many mutations of the phenylalanine
hyperphenylalaninemia. hydroxy lase gene have been identified (missense,
nonsense, insertions, deletions, and duplications)
leading to PKU or non-PKU hyperphenylalaninemia.
Phenylketonuria (PKU) The incidence of classic PKU is about I in
Deficiency of phenylalanine hydroxylase, 10,000-
tetrahydrobiopterin, or dihydropteridine reductase 20,000 live births and exhibits considerable
results in phenylketonuria (PKU), an autosomal geographic variation (the incidence in Ireland is 1 in
recessive trait. Be- cause phenylalanine accumulates 4000, whereas the condition is rare among blacks
in tissues and plasma (hyperphenylalaninemia), it is and Asians). About
metabolized by alternative pathways and abnormal 2% of hyperphenylalaninemic infants have a
amounts of phenylpyruvate ap- pear in urine (Figure deficiency of biopterin or biopterin reductase. The
17-22). Phenylalanine hydroxylase deficiency may be most important clinical presentation is mental
complete (classic PKU, type I) or partial impairment. Diagnosis can be made early in the
359
neonatal period by measurement of phenylalanine Metabolism
concentration in blood collected from
360
Metabolism
H tinuation throughout the first decade, or for life, may be

I
HI H�¥�x;
l�s
YNH,
NH j
necessary.
Treatment of biopterin and biopterin reductase defi-
ciency consists not only of regulating the blood levels of
H,C-C-C N
phenylalanine but of supplying the missing form of coen-
bH bH i O zyme and the precursors of neurotransmitters, namely,
A-group dihydroxyphenylalanine and 5-hydroxytryptophan, along
5,6, 7,8· Tetrahydrobiopterin with a compound that inhibits peripheral aromatic de-
carboxylation. This compound is necessary because the
amine products do not cross the blood-brain barrier.
Successfully treated females who have reached repro-
ductive age may expose their offspring (who are obli-
gate heterozygotes) to abnormal embryonic and fetal de-
velopment. These effects include spontaneous abortion,
Para-quinonoid Ortho-quinonoid microcephaly, congenital heart disease, and intrauterine
dihydropteridine dihydropteridine
growth retardation, and they correlate with the plasma
"Quinonoid" dihydrobiopterins level of phenylalanine of the pregnant mother. Thus,
H
��Nx;YNH,
I reinstitution of a low-phenylalanine diet during pre-
and postconception periods may be necessary. The diet
H� I NH
@ N
should also restrict intake of phenylalanine-containing
substances, such as the synthetic sweetener aspartame (L-
0 aspartyl-L-phenylalanyl methyl ester). Because defective
7,8-Dihydrobiopterin
myelination occurs in the brain in PKU, there is an in-
creased incidence of epileptic seizures and abnormal elec-
FIGURE 17-21 troencephalograms are common. The biochemical basis
Structures of biopterin derivatives.
for the severe mental impairment is not understood. One
factor may be inhibition of glutamate decarboxylase by
a heel prick onto filter paper. Treatment of phenylalanine phenylpyruvate and phenylacetate:
hydroxylase deficiency consists of a diet low in pheny-
o�
lalanine but which maintains normal nutrition. This diet
is effective in preventing mental retardation, and its con-
Glutamate decarboxylase
co,
I '
O
Transaminase
r
-00C-(CH2b-NH3 +
NH+ L-
CH-CH-coo- (pyridoxal pho�hate) 'I � CH-c-
coo- 1
Phenylpyruvate
Glutamate y-Aminobutyrate
a-Ketoglutarate Glutamate -
2
Phenylalanine
y--NADH + H+
l'-NAD+ The substrate is an excitatory neurotransmitter and the

product an inhibitory one in the central nervous system.·
OCH,COO- co, OcH,-f- Abnormal indole derivatives in the urine and low
cooH levels of serotonin (a product of tryptophan metabolism) in
Phenylacetate
OH blood and brain point to a defect in tryptophan metabolism
In they--Glutamine Phenyllactate
liver!'- H,0
in PKU. 5-Hydroxytryptophan decarboxylase, which
catalyzes the conversion of 5-hydroxytryptophan to
��Too
0 -
- CH-C-N-CH
' I
CH,
I
serotonin, is inhibited in vitro by some of the metabolites
of phenylalanine. Phenylalanine hydroxylase is similar to
the enzyme that catalyzes the hydroxylation of tryptophan
r to 5-hydroxytryptophan, a precursor of serotonin. In vitro,
361
CONH, Metabolism
Phenylacetylglutamine phenylalanine is also found to inhibit the hydroxylation of
FIGURE 17-22 tryptophan. The mental defects associated with PKU may
Formation of metabolites of phenylalanine that accumulate in abnormal be caused by decreased production of serotonin. High
amounts and are excreted in phenylketonuria. phenylalanine levels may disturb the transport of amino
360
a
acids into cells. Variations in the clinical manifestations of c�
PKU may reflect differences in this disturbance. Defects H-c-coo- Tyrosinase
r \
HOa I qi,\
H-C-COO
_
in pigmentation of skin and hair (light skin and hair) HO H,/ [OJ H,0 HO � +H,/
may be caused by interference of melanin formation by
Tyrosine 3,4-Dihydroxyphenylalanine
phenylalanine and its metabolites and also by lack of (dopa)
tyrosine.
[OJ� Tyrosinase
H,0-1
Melanin 0
0
Melanin is an insoluble, high-molecular-weight poly- � - - - YYac\-coo-
mer of 5,6-dihydroxyindole, which is synthesized from
tyrosine (Figure 17-23). It is produced by pigment cells
o��coo-
H
oA/ +�/
Dopa quinone
(melanocytes) in cytoplasmic organelles (melanosomes). Dopachrome
In the epidermis, melanocytes are associated with lzn'+

keratinocytes, which contain melanosomes supplied by
melanocytes via dendritic processes. Color variation in
human skin reflects the amount of melanin synthesized
in melanosomes. Melanin synthesis is apparently under
hormonal and neural regulation. HOMN,)
co,-1
HOY'r-i
ow
0
# I
-
H H
atalyzed by tyrosinase, a copper-containing oxidase, Cysteine -

l
The first two steps in the synthesis of melanin are 5.6-Dihydroxy indole lndole-5 ,6- qui none
c
which converts tyrosine to dopaquinone. All subse- Trichromes; -
quent reactions presumably occur through nonenzymatic I Polymerization [
l
auto-oxidation, in the presence of zinc, with forma- I
Pheomelanins H
tion of the black to brown pigment eumelanin. The N 0
yellow to reddish brown, high-molecular-weight poly-
mer known as pheomelanin and the low-molecular-weight 0
trichromes result from addition of cysteine to dopaquinone
and further modification of the products. Pheome-
lanins and trichromes are primarily present in hair and
feathers.
H
N
Abnormalities of Tyrosine Metabolism
Hepatic cytosolic tyrosine aminotransferase (tyrosine
transaminase) deficiency produces tyrosinemia type II, Eumelanins
an autosomal recessive trait marked by hypertyrosine- FIGURE 17-23
mia and tyrosinuria. Clinical manifestations may include Biosynthesis of
corneal erosions and plaques, inflammation (from intracel- melanins.
lular crystallization of tyrosine), and mental retardation.
Low-tyrosine and low-phenylalanine diets are beneficial. homogentisic acid. The biochemical lesion is homogen-
Tyrosinosis is presumably due to fumarylacetoacetate tisic acid oxidase deficiency. Clinical features include
hydrolase deficiency and has a high prevalence in the pigmentation of cartilage and other connective tissues
French-Canadian population of Quebec. It is associ- (ochronosis) later in life from deposition of oxidized ho-
ated with abnormal liver function, renal tubular dys- mogentisic acid. Patients nearly always develop arthritis
function, anemia, and vitamin D-resistant rickets. Tran- in later years, but the relationship between pigment depo-
sient tyrosinemia of the newborn, particularly in prema- sition and arthritis is not understood.
ture infants, is the most common form of tyrosinemia in Lack of melanin production (hypomelanosis) gives rise
infancy. to several hereditary disorders collectively known as al-
Alcaptonuria is a rare metabolic hereditary disease in binism. Some forms result from deficiency of tyrosinase.
which homogentisic acid is eliminated in urine, which The inheritance pattern of albinism varies with type. Af-
darkens upon exposure to air owing to oxidation of fected individuals have increased susceptibility to various
361
Tryptophan pyrrolase
o,
N·Form�
lkyz�enine
mamidase
Formate -One-carbon pool
0
II _
C-CH- CH-COO
CX
Kynurenine , I
hydroxylase
N�
0,, NADPH + H+
NH,
Kynurenine
3-Hydroxykynurenine
Kynureninase�H,O
(pyridoxal
phosphate) Alanine
coo- �
-- -NAO+
NH+3
YN�coo-
OH
OH
Xanthurenate
3-Hydroxyanthranilate (excreted in urine)
FIGURE 17-24
Pathway for the synthesis of NAD from tryptophan.
types of carcinoma (from the effect of solar radiation on ate precursor of NAO. In deficiency of pyridoxal phos-
DNA). When the eyes are involved, photophobia, sub- phate, 3-hydroxykynurenine accumulates and is converted
normal visual acuity, strabismus, and nystagmus may be to xanthurenate, which is excreted in urine. Thus, vitamin
present. B6 deficiency can be diagnosed by measurement of uri-
nary xanthurenate after administration of a standard dose
of tryptophan (tryptophan load test).
Tryptophan 5-Hydroxytryptamine (serotonin) is found in ente-
rochromaffin cells, brain, and platelets. In the former two,
Tryptophan is an essential amino acid involved in it is produced from tryptophan, whereas in platelets, sero-
synthesis of several important compounds. Nicotinic tonin is taken up from plasma. Synthesis involves hydrox-
acid (amide), a vitamin required in the synthesis of ylation of tryptophan by tryptophan 5-hydroxylase and
NAD+ and NADP+, can be synthesized from tryptophan decarboxylation by aromatic L-arnino acid decarboxylase
(Figure 17-24). About 60 mg oftryptophan can give rise to (Figure 17-25). Hydroxylation is the rate-limiting reac-
1 mg of nicotinamide. The synthesis begins with conver- tion, is analogous to that of phenylalanine, and requires
sion of tryptophan to N-formylkynurenine by tryptophan molecular oxygen and tetrahydrobiopterin. Serotonin is a
pyrrolase, an inducible iron-porphyrin enzyme of liver. powerful vasoconstrictor and stimulator of smooth mus-
N-Formylkynurenine is converted to kynurenine by re- cle contraction. In the brain it is a neurotransmitter, and
moval of formate, which enters the one-carbon pool. in the pineal gland it serves as a precursor of melatonin.
Kynurenine is hydroxylated to 3-hydroxykynurenine, Synthesis of melatonin requires N-acetylation of sero-
which is converted to 3-hydroxyanthranilate, catalyzed tonin, followed by methylation (Figure 17-26). The role
by kynureninase, a pyridoxal phosphate-dependent en- of melatonin in humans is not understood; in frogs, it
zyme. 3-Hydroxyanthranilate is then converted by a lightens the color of skin melanocytes and blocks the
series of reactions to nicotinamide ribotide, the immedi- action of melanocyte-stimulating hormone (MSH) and
362 CHAPTER 17 Protein and Amino Acid Metabolism
NH.. adrenocorticotropic hormone (ACTH). In rats, melatonin
I •
�CH;-<Olt--C00- regulates the breeding cycle, and its secretion is increased
by exposure to light via adrenergic stimulation of the
pineal gland.
H
1 N-Acetyltransferase is activated by increased concen-
Tryptophan trations of cytosolic cyclic AMP and Ca2+ that is medi-
0,, tetrahydrobiopterin� ated by the activation of adrenergic receptors of the pineal
H,O, dihydrobiopterin _,.. Tryptophan-5-hydroxylase
1 gland. In humans, melatonin synthesis and its release fol-
NH+
HO I • lows a circadian rhythm which is stimulated by dark-
�CH,�Cft--C00-
ness and inhibited by light. The blood levels of melatonin
increase by passive diffusion from the central nervous sys-
tem after the onset of darkness, reaching a peak value dur-
1 ing the middle of the night and declining during the second
H
5-Hydroxytryptophan half of the night. Melatonin secretion is also regulated en-
Aromatic L -amino acid dogenously by signals from the suprachiasmatic nucleus.
CO, decarboxylase Since melatonin's peak concentration in plasma coincides
(pyridoxal phosphate)
� with sleep, exogenous administration of the hormone can
affect the circadian rhythm. Melatonin supplementation
HO�CH;--CH,NH: has been used to ameliorate subjective and objective symp-
�N) toms of jet lag caused by travel across time zones. At high
levels, melatonin promotes sleep. Short-term and long-
H
I
term biological effects of melatonin supplementation have
5-Hydroxytryptamine
(serotonin) yet to be determined.
Serotonin is degraded to 5-hydroxyindoleacetic acid
FIGURE 17-25
Biosynthesis of serotonin from tryptophan.
(5-HIAA) by monoamine oxidase and aldehyde dehy-
drogenase acting in sequence. 5-HIAA is excreted in
the urine. Its excretion is markedly increased in subjects
with carcinoid tumor (found most frequently in the gas-
trointestinal tract and lungs). Carcinoid tumors are fre-
HO�CH- CH-NH+
2
quently indolent and asymptomatic; however, a signifi-
� •• ) Serot�nin ' cant number of these tumors can manifest as metabolic
7
H
problems. Although increased production of serotonin
is a characteristic feature of the carcinoid tumor, cells
Acetyl-CoA�5-Hydroxytryptamine- also synthesize other substances. These include kinins,
CoASH N-acetyltransferase prostaglandins, substance P, gastrin, somatostatin, corti-
0 cotropin, and neuron-specific enolase. Carcinoid tumor
U..)
H� CH-
2 CH-NH��-CH
N-Ac;tyl·5-hydroxytryp�amine
cells are known as enterochromaffin cells because they
stain with potassium dichromate and also are known as
argentaffin cells because they take up and reduce silver
7
H salts. The symptoms of the carcinoid tumor are due to syn-
S-Adenosylmethionine�
ergistic biochemical interactions between serotonin and
. Hydroxyindole methyltransferase the above-mentioned active metabolites. Characteristics
S-Adenosy lhomocyste me
0 of the carcinoid syndrome are flushing, diarrhea, wheez-
H,COO= r
� I I CH,CH,NH-C-CH,
II ing, heart valve dysfunction, and pellagra. Lifestyle con-
ditions that precipitate symptoms include intake of alco-
hol or spicy foods, and strenuous exercise. The treatment
' N options for the carcinoid syndrome are multidisciplinary,
I
H including lifestyle changes, inhibitors of serotonin re-
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FIGURE 17-26 logues, hepatic artery embolization, chemotherapy, and
Biosynthesis of melatonin from serotonin. surgery.
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340

17.2 Metabolism of Ammonia

Ammonia (at physiological pH, 98.5% exists as NHt),

NH; 2ATP 2ADP +

acids produced in excess compete for CoA-SH for

NAG binding changes the conformation and subunit

Formation of Citrulline Formation ofArginine and Fumarate

the enzymes discussed above, an exogenous supply of Hyperammonemias

O-!-s-coA + AMP2- + pp,3- O CH3-C-SCoA + H3 •N-CH

The excretion of phenylacetylglutamine produces loss of

The overall reaction is: interferon-y ), or bacterial lipopolysaccharides can

the vascular endothelium, agonists such as

system (Chapter 30 ). The NO produced in the vascular ,l,,,�.Jl H2

muscle cells. Organic nitrates used in the management of

pulmonary vasodilation. This property of NO has been 0 o,s,Nl

rea------ '\_ r ------Pyruvat�

Bile acid conjugates / I \ <, l /

in Figure 17-9) is used orally in the therapy of some

NHt + C02 + NADH + N5,N10-methylene-FH4

Glyoxalate can be transaminated to glycine, reduced

Arginine Glycine Guanidinoacetate Ornithine

The glyoxylate that accumulates is converted

Carboligase Thiamine pyrophosphate

This kinase is a dimer of M and B (M = muscle, B

Decarboxylation of omithine to putrescine by omithine decarboxylase. Omithine aminotransferase deficiency is

!,-- Glutamate t:,. -Pyrroline-

of collagen (Chapter 25). Hydroxyproline released by N5-methyltetrahydrofolate and is unavailable as a carrier

carboxylate dehydrogenase is deficient.

late binders to the vitamins. High urinary levels of Figlu

of histamine). Its effect on secretion of hydrochloric acid Leucme. isoleuone. and

F,-ro-m-e_ucl.. ,. n� j From 1soleucinej

The o-keto acids, by oxidative decarboxylation, yield FIGURE 17-14

Hypoglycin produces hypoglycemia and metabolic aci-

acids is necessary to monitor the degree of dietary restric-

aciduria (sweaty feet), 3-methylcrotonylglycinuria ( cat's 1. Guanldinoacetic acid

Betaine (an acid) is obtained from oxidation of choline

NAO• lSetaine aldehyde

Cysteine carboxylic acid group. Taurine is conjugated with bile

o-w-o ,��rase -o-M-o-[-o-p

the methylation of homocysteine to methionine remethylation or transsulfuration pathways. Individuals

ydrofolate reductase (DR)

Methylene FH4 Methylation

o-v- NH+ / � NH;

CH;-CH-coo- )> hydroxylase

Phenylalanine biopterin dihydrobiopterin Tyrosine

Hydroxylation, shift� H�roxyphenylpyruvic

H tinuation throughout the first decade, or for life, may be

l'-NAD+ The substrate is an excitatory neurotransmitter and the

In the epidermis, melanocytes are associated with lzn'+

atalyzed by tyrosinase, a copper-containing oxidase, Cysteine -

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