1-Introduction To Enzymes
1-Introduction To Enzymes
1-Introduction To Enzymes
Enzymes activity Ex. Drugs & Ex. Cofactors (inorganic substances) such
as Ca++, Mg++,Mn++,Cu++ and Zn++ {can be
Poisons.
removed by dialysis
And riboflavin}.
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Enzymes can be classified according to their structure to:
Properties of enzymes:-
3-reactants can work without enzymes, but enzymes fasten the reaction.
4-reactants must have sufficient energy for the reaction to proceed, which
is not needed for all the reaction molecules.
10-like all proteins, made of long linear chain of a.a. that fold to produce
3D. Structure. Each with specific sequence and properties.
12-some enzymes are larger than its substrate and only small portion of
enzyme {3-4 amino acids} is involved in catalysis, this portion called the
active site.
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14-they lower the barriers that normally prevent the chemical reaction
from occurring or slow them down by decreasing the required activation
energy. Thus, in presence of enzyme the reaction proceed in a faster rate.
15-they’re usually named with the suffix –ase at the end of their substrate
name. EX1.sucrose→sucrase.
EX2.lactose→lactase.
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2-SPECIFICITY
The ability of an enzyme to catalyze the specific reaction and no others is
perhaps its most significant property.
However specificity varies from one enzyme to another, so we have the
following types of enzyme specificity.
1) Absolute specificity
Catalyze only one substrate not the other.
Ex1. Urase acts only on urea
Ex2.Fumerase acts only on fumrate
⁻OOC.CH=CH.COO⁻ FUMERASE ⁻OOC.CHOH.CH₂.COO⁻
L.Fumerate H2O L.malate
2) Broad specificity
The enzyme is specific to one type of bond possessed by a group of
chemically related substances.
Ex1.Pancreatic lipase hydrolysis alpha ester bonds (at position 1 and 3) of
triglycerides.
Ex2. Salivary amylase attack alpha 1:4 glucosidic bonds of starch.
Ex3.Polypeptidase.
3) Group specificity
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A- Act on specific chemical group such as
➢ glycosidases on glycosides , pepsin and trypsin on peptide bonds
and esterases on esters.
➢ ii-Lytic enzymes exhibit a higher order of group specificity; e.g
carboxypeptidases and aminopeptidases split off amino acids one at
a time from the carboxy or amino terminal end of polypeptide chains.
B-Linkage specificity is related to group specificity and it act on a
particular type of chemical compound regardless of the rest of the
molecular structure.
3-NAMING OF ENZYMES
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Among the first identified and named were the digestive enzyme that
catalyzes the hydrolysis of the glycoside, ester and amide bond of
carbohydrates, lipids and proteins.
2. Enzymes were named for the substrates on which they acted by adding
the suffix -ase to the substrate such as proteinase , lipase.
3. Enzymes given the name of chemical reaction such as oxidases ,
decarboxylases and dehydrogenases.
4. The same enzyme being identified with two different names. One at the
normal physiological reaction and other at artifical conditions.
Ex1. Glucose isomerase industrially to convert glucose into the
sweetner fructose.
EX2. Xylose isomerase in vivo.
5. Naming depends on the types and mechanisms of chemical reactions.
a) Reactions and enzymes that catalyze them divided into 6 classes each
with 4-13 subclasses.
b) The enzyme names has 2 parts , the first is the substrate, second the
ending –ase
c) Each enzyme has a systemic code number (E.C). This number
characterized the reaction type. The Class,Subclass,Sub subclass then
the specific name.
E.C 2.7.1.1 denotes
Class 2 (transferase)
Subclass 7 (transfer of phosphate)
Sub subclass 1 (an alcohol function as the phosphate acceptor)
The enzyme name (hexokinase)
E.C 3.4.11.4 (tripeptide aminopeptidases
Class 3 (hydrolases)
Subclass 4 (hydrolases act on peptide bonds)
Sub subclass 11 (hydrolases that cleave off the amino-terminal aminoacid
from a polypeptide)
The enzymes are those that cleave off the amino-terminal end from a
tripeptide.
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4-CLASSIFICATION OF ENZYMES
1) Oxidoreductases
Are biological oxidation and reduction between 2 substrate and there for
are essential in the process metabolism energy releasing, respiration and
fermentation.
3) Hydrolases
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Ex. phosphatase catalyze dephosphorylation {removal of phosphate
group}
4) Lyases
5) Isomerases
Ex. Rotamase and protein disulfide isomerase which can assist apeptide
chain to fold into a correct 3D structure.
b) D-glucose L-glucose
6) Ligases
Catalyze the reaction which joins two molecules and formation of different
bonds by condensation reaction coupled to ATP cleavage; C-C, C-N, C-S
and C-O.
Coenzymes-5
Definition
Some enzymes are purely protein in nature and depend for activity on
their structure, while certain enzymes requires for their function, one or
more non-protein part, these are termed as Coenzymes, Cofactors or
prosthetic groups.
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If such a compound is firmly attached to the enzyme protein then it’s
called Coenzyme.
Properties of coenzymes:
➢ They are organic molecules that cooperate in the catalytic action of the
enzyme.
Ex. NADH, NADPH & ATP (that are released from the enzyme’s active site
during the reaction.
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They’re water soluble and diffuse freely away from the enzyme after the
oxidation or reduction reaction is completed to join another
dehydrogenase reaction.
The general form of reactions involving NAD⁺ is:
RH2+ NAD⁺↔NADH + H⁺
4. Thiamine pyrophosphate
Thiamine (B₁) is the precursor of thiamine pyrophosphate, is the
coenzyme for some important oxidation-decarboxylation reactions
Ex. Pyruvate dehydrogenase and Oxoglutarate dehyrogenase.
5. Coenzyme A
Complex group contains sulphahydryl (--SH) group.
This group reacts with carboxyl group.
6. Biotin coenzyme
Coenzyme for carboxylation reactions.
Ex. Pyruvate carboxylase.
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6-Mechanism of enzymes
Activation energy:
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Internal dynamics is the movement of parts of the enzyme’s structure, for
example:
• Individual amino acid.
• Group of amino acids.
• Entire protein domain.
7-Enzymes Regulation
Many of enzyme reactions occur in cells involving synthesis, degradation
or interconversion.These reactions regulate by 2 ways
Ex1. Enzymes in the liver called cytochrome P450 oxidases, which are
important in drug metabolism. Induction or inhibition of these enzymes
can cause drug interactions.
Ex2.If a new substrate is made available to the cell; it may induce the
synthesis of the enzymes needed to cope with it.
B) Enzyme repression
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The repressors which may be a product or hormone affect DNA to stop
synthesis of specific protein which is an enzyme.
Ex.If ample quantities of an amino acid are already available to the cell
from its extracellular fluid; synthesis of the enzymes that would enable
the cell to produce that amino acid for itself is shut down.
When the regulator binds to its site, it alters the shape of the enzyme so
that its activity is changed.
The end product(s) of a metabolic pathway are often inhibitors for one of
the first enzymes of the pathway (usually the first irreversible step, called
committed step), thus regulating the amount of end product made by the
pathways.
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The amount of the end product produced is regulated by its own
concentration.
This helps allocate materials and energy economically, and prevents the
manufacture of excess end products.
The enzymes are produced in inactive form and when it reaches the site
of reaction it is activated.
This stops the enzyme from digesting the pancreas or other tissues before
it enters the gut.
D) Regulatory protein
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A Ca+2 bind to calmodulin induce a co-formational change that convert it
from inactive to active form.
Activated calmodulin bind to many enzymes and other proteins in the cell
and modify their activity either by stimulation or inhibition.
E) Covalent modification
Enzyme rate
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Conc. Of sub. And enzyme
2) Temperature:
When the temperature increase the rate of the reaction will increase even
without enzyme but with low rate .But when add enzyme it accelerates
the reaction, because it removes barriers.
Activation velocity
Plato
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Inactivation
Activation temperature
3)PH:
Enzyme PH Optimum
Enzyme Inhibition-9
Enzymes can be inhibited by many substances and reduce the initial
velocity of such enzymes. Accordingly there are 2 types of enzymes
inhibition.
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1) Irreversible inhibitors
React with the enzyme and form a covalent adducts with the protein. The
inactivation is irreversible. These compounds include eflornithine a drug
used to treat the parasitic disease sleeping sickness. Penicillin and Aspirin
also act in this manner. With these drugs, the compound is bound in the
active site and the enzyme then converts the inhibitor into an activated
form that reacts irreversibly with one or more amino acid residues.
2) Reversible inhibitors
Reversible inhibitors bind and dissociate with their enzyme in equilibrium
process. It can be classified as
a) Competitive inhibition
b) Non competitive inhibition
c) Uncompetitive inhibition
d) Mixed inhibition
A) Competitive inhibition
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B) Non competitive inhibition
Can bind to the enzyme at the same time as the substrate, i.e. they never
bind to the active site. Both the EI and EIS complexes are enzymatically
inactive. Because the inhibitor cannot be driven from the enzyme by
higher substrate concentration (in contrast to competitive inhibition), the
apparent Vmax changes. But because the substrate can still bind to the
enzyme, the Km stays the same.
Occur in cells when specific molecule bind to an enzyme site other than
the active site called Allosteric site causing a shift in the enzyme
configuration which prevent the substrate from binding to the active site.
C) Uncompetitive inhibition
D) Mixed inhibition
This type of inhibition resembles the non-competitive, except that the EIS-
complex has residual enzymatic activity.
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In many organisms inhibitors may act as part of a feedback mechanism. If
an enzyme produces too much of one substance in the organism, that
substance may act as an inhibitor for the enzyme at the beginning of the
pathway that produces it, causing production of the substance to slow
down or stop when there is sufficient amount. This is a form of negative
feedback. Enzymes which are subject to this form of regulation are often
multimeric and have Allosteric binding sites for regulatory substances.
Their substrate/velocity plots are not hyperbolar, but sigmoidal (S-
shaped).
Uses of inhibitors:
Since inhibitors modulate the function of enzymes they are often used as
drugs and poisons.
Enzyme units:
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Enzyme units are expressed in micromoles (10-6 mol), nanomoles (10-9
mol) or picomoles (10-12 mol) of substrate reacting or product produced 1
minute.
Isozyme
It is the presence of enzyme in different forms but all have the same
reaction differ from each other in some chemical or physical properties.
m-dehydrogenase
EX1.Malate oxaloacetate
Proenzymes
Many proteins are manufactured and secreted from cells in the form of
inactive precursor or proteins known as "proproteins". When the proteins
are enzymes, they are termed proenzymes or zymogens.
Ex1.Insulin proinsulin
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Ex2.Pepsin pepsinogen
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LDH 1 – Found in heart and red-blood cells and is 17% – 27% of the normal
serum total.
LDH 2 – Found in heart and red-blood cells and is 27% – 37% of the normal
serum total.
LDH 3 – Found in a variety of organs and is 18% – 25% of the normal
serum total.
LDH 4 – Found in a variety of organs and is 3% – 8% of the normal serum
total.
LDH 5 – Found in liver and skeletal muscle and is 0% – 5% of the normal
serum total.
Following a myocardial infarct the serum levels of LDH rise within 24-48
hours reaching a peak by 2–3 days and return to normal in 5-10 days.
Especially diagnostic is a comparison of the LDH-1/LDH-2 ratio. Normally,
this ration is less than 1. A reversal of this ration is referred to as a
“flipped LDH”. Following an acute myocardial infarct the flipped LDH ratio
will appear in 12–24 hours and is definitely present by 48 hours in over
80% of patients. Also important is the fact that persons suffering chest
pain due to angina only will not likely have altered LDH levels.
Diagnostic and prognostic value of specific enzymes
The determination of the activity of the following enzymes can provide the
physician with valuable diagnostic evidence.
1-Lipase
The plasma lipase level may be low in liver disease, vitamin A deficiency,
some malignancies and diabetes mellitus. It may be elevated in acute
pancreatitis and pancreatic carcinoma.
2-Amylase
The plasma amylase may below in liver disease and increased in acute
pancreatitis and diabetes mellitus.
3-Trypsin
Elevated level of trypsin in plasma occurs during acute pancreatic
disease. The elevation is more sensitive and considers indicator of
pancreatic disease than plasma amylase or lipase.
4-Cholinesterase
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In general, low levels are found in patients ill with liver disease
malnutrition, acute infection disease and anemia. High levels occur in
nephritic syndrome.
Since the content of cholinesterase in young red blood cells is
considerably higher than in the adult red blood cells, the cholinesterase
titer of erythrocytes in peripheral blood gives an indication of
hematopoietic activity.
5-Alkaline phosphatase
The level of enzymes capable of catalyzing the hydrolysis of various
phosphate esterase at alkaline PH (alkaline phosphatase activity) may be
increased in hyper parathyrodisim and osteoblastic sarcoma.
Isozymes of alkaline phosphatase are present in body fluid. These include
specific isozymes originating from bone, liver, placenta and intestine.
Measurement of specific alkaline phosphatase isozymes may therefore
improve the diagnostic value of this test.
6. Acid phosphatase
The level of enzymes capable of catalyzing the hydrolysis of various
phosphate esters at acidic PH (acid phosphatase activity) may be elevated
in the metastatic prostatic carcinoma.
7. Transaminases
Two transaminases are of clinical interest
a) Glutamic oxaloacetic transaminases (GOT) which catalyzes the transfer
of the amino group of aspartic acid to alpha-ketoglutaric acid, forming
glutamic and oxalo acetic acids.
b) Glutamic pyruvic transaminase (GPT) transfers the amino group of
alanine to alpha-ketoglutaric acid, forming glutamic and pyruvic acids.
Liver tissue rich in both transaminases are elevated in sera than GOT.
While both transaminases are elevated in sera of patients with acute
hepatic disease, GPT, which is only elevated by cardiac necrosis, is a
more specific indicator of liver damage.
8. Lactate dehydrogenase
In myocardial infarction, the concentration of serum lactate
dehydrogenase (LDH) rises within 24hrs after the infract and return to the
normal range within 5-6 days. High levels of LDH also occur in patients
with acute and chronic leukemia in relapse.
9. LDH isozymes
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Cardiac muscle contains a preponderance of LDH-1.
Measurements of the serum isozyme pattern following myocardial
infarction is a more sensitive and lasting indication of myocardial necrosis
than the simple measurements of the total serum or plasma LDH activity.
Y
oussra Sayed Othman.
M
arwa Mohamed Amin.
Di
na Gamal Tawfik.
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