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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 mu- cosa 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 in- volves deamination with
production of ammonia, which is removed by formation
of glutamine. N-acetylaspartate oc- curs in high levels in
the brain but its function is not known. It is synthesized
from acetyl-CoA and aspartic acid cat- alyzed 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 usu- ally of
Eastern European Jewish heritage.
Urea Synthesis
Ammonia contained in the blood flowing through the
hep- atic lobule is removed by the hepatocytes and
converted into urea. Periportal hepatocytes are the
predominant sites of urea formation. Any ammonia that
342
SECTION 11.2 Metabolism of Ammonia CHAPTER 17Protein and Amino Acid 341
Metabolism
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
NAG.K'.Mg2'
NH,++ HC0 + 2ATP4 3- -
0
II
H2N-C-OPO/- + 2ADP3- + p/- + 2H +
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.
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'
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 en- dothelial 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
SECTION Metabolism of Some Individual Amino Acids CHAPTER 11Protein and Amino Acid 345
Metabolism
NH3+
Citrulline
Signal Transduction of NO
The NOS activity is inhibited by NG-substituted
ana- logues of arginine, such as NG-nitroarginine NO is lipophilic and diffuses readily across cell
and NG- monomethyl-L-arginine. mem- branes. It interacts with molecules in the target
cells pro- ducing various biological effects. One
mechanism of ac- tion of NO is stimulation of
Isoforms (Also Known as Isozymes)
guanylate cyclase, which catalyzes the formation of
of Nitric Oxide Synthase
cyclic guanosine monophos- phate (cGMP) from
There are three major isoforms of Nitric GTP, resulting in increased intracel- lular 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 activa- tion 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)
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
·�lntnx
"One-carbon... + NH;�;lyc\
' Serine� Ethanolamine- Cho::: cycle-co,
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
physiologi- cal 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. path- ways 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 caver- nosum. This compound (structure shown pyridoxal phosphate
leading
to increased production of cGMP. Smooth muscle Signal transduction of NO by cGMP-
relax- ation 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+ contain- ing 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 cardiovas- cular system.
348
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 Protein and Amino Acid 347
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 reac- tions
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
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 guanidinoac- etate) requires transamidination,
� i.e., transfer of a guani- dine group from arginine to
glycine glycine, to form guanidinoac- etate (glycocyamine) by
CHO-coo- + NHt + mitochondrial arginine-glycine amidinotransferase
H202 NH2
glyoxalate
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
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 car- diac 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 crea- tine 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
supplementa- tion 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
inter- mediate 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 glu- tamate, 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 mus- cle 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 transam- inase, 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 in- volved 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 crea- tine
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
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 350
Protein and Amino Acid Metabolism
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
pro- line (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
transami- nated to ornithine or reduced to glutamate
(Figure 17-12).
350
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 351
Protein and Amino Acid Metabolism
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�-.
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
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 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
:
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
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
FAO FAOH2
(CH3h •N-CH2-CH20H :::,,, ,/ • (CH3h •N-CH2-CHO
Choline o>cidase
Choline Betaine aldehyde
S-Adenosylmethionine (CH3l3N-CH,-Coo-
(active methionine) Betaine
354
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 17 Protein and Amino Acid 355
Metabolism
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).
Folate (F)
2 NADPH+2H+
Tetrahydrofolate (FH4)
Methyl
Serine S- acceptor
adenosylmethionine s
Serine
hydroxymethyl
transferase
(pyridoxal phosphate) Methylate
d acceptor
Glycine
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
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 Protein and Amino Acid 357
Metabolism
synthesis of a variety of important compounds, includ-
ing thyroxine, melanin, norepinephrine, and epinephrine
356
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 11 Protein and Amino Acid 357
Metabolism
DIET
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
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 17Protein and Amino Acid 358
NH+
I • -0 , NH+
Metabolism
O
Q. Phenylalanine H,O
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
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
hyper- phenylalaninemia. hy- droxy 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-
tetrahy- drobiopterin, 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
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 17Protein and Amino Acid 359
neonatal period by measurement of phenylalanine Metabolism
concentration in blood collected from
360
SECTION 17.3 Metabolism of Some Individual Amino Acids CHAPTER 17Protein and Amino Acid 360
Metabolism
��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+
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
HOY'r-i
ow
0
# I
-
H H
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-
Melatonin (N-acetyl-5-methoxytryptamine) lease and serotonin receptor antagonists, somatostatin ana-
FIGURE 17-26 logues, hepatic artery embolization, chemotherapy, and
Biosynthesis of melatonin from serotonin. surgery.
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