Michaelis-Menten equation

Km = [S] at Vmax (1/2 of enzyme bound to S);

Vmax = velocity where all of the enzyme is bound to substrate
(enzyme is saturated with S).
Assume that [S]>>[E]; k+2<< k+1; k+2<< k-1
V formation of ES = V degradation of ES

k+1([E] - [ES]) [S] = k-1 [ES] + k+2 [ES]

 [E] [S] k+2[ES] [S]
[ES]= V=
Km + [S] Km + [S]

Vmax [S]
k-1+ K+2
Km= V=
k+1 Km + [S]
From this kinetic scheme, a relationship
can be derived for the rate or velocity of
the reaction:
Michaelis-Menten Equation

gives hyperbolic curve

Km = [S], where the velocity V = Vmax/2,
is called the Michaelis constant.

- Km is in units of concentration
- Km good estimate for the optimum
concentration of substrate.
A plot of

is not accurate enough to derive good

Km & Vmax.
Computer analysis is done.
Reciprocal Plot
A double reciprocal plot or
Lineweaver-Burk plot is linear and more
eye-appealing for presentation.
mathematically = linear transformation
Lineweaver-Burke equation
 Effect of pH and
temperature on the velocity
Lines intersect at
the y-axis
 Inhibition of enzyme activity
COMPETITIVE INHIBITION
S and I are structure analogs;
Both act in the active site;
ES or EI complex is formed;
The degree of inhibition depends on the ratio [S] / [I].
 Inhibition of enzyme activity
NONCOMPETITIVE INHIBITION
S and I are different;
I act in the allosteric site;
ES, EI and ESI complexes are formed;
The degree of inhibition depends only on the [I].
Purine and pyrimidine analogs as
anticancer and antiviral agents
 Treatment of hypercholesterolemia with
statins competitive inhibitors of HMG CoA
 Antifolate drugs - affect the folate metabolism
Methotrexate - competitive inhibitor of dihydrofolate reductase. Inhibit
cell division and is used in the treatment of some hematological (or other)
malignancies.

Sulfonamides - competitive inhibitors of p-aminobenzoic acid. Used to

treat bacterial infections. When bacteria try to incorporate sulfonamides into the
molecule of THF, the resulting molecule possesses a non-functional cofactor.
Zymogen activation by proleolytic cleavage
Feedback inhibition
Feedback inhibition - Regulation of purine synthesis de novo
Treatment of gout with allopurinol
Competitive inhibitor of xanthine oxidase
Feedback inhibition - Regulation of pyrimidine synthesis
Regulation of enzyme activity with phosphorylation /
dephosphorylation
Regulation of TAG degradation and synthesis in adipose tissue

Fatty acids synthesis

Malonyl CoA

Acetyl CoA

Acetyl CoA Acetyl CoA

carboxylase carboxylase
Phosphatases
(INSULIN)
Phosphatases
(INSULIN)
 Clinical significance of enzymes
Creatine kinase

Dimer
Subunit M (Muscle)
Subunit B (Brain)

Three isoforms:
MM, MB, BB
 Clinical significance of enzymes
Lactate dehydrogenase

Tetramer
Subunit H (Heart)
Subunit M (Muscle)

Five isoforms:
HHHH (LDH1)
HHHM (LDH2)
HHMM (LDH3)
HMMM (LDH4)
MMMM (LDH5)
Clinical significance of enzymes at
myocardial infarction