Catalytic Proteins- Enzymes

Part 2

Kinetics and Enzyme Regulation

 Michaelis Menten Model
kcat

E+S Rapid equilibrium

ES Rate limiting step
E+P

The maximum velocity that a reaction could reach in

constant enzyme concentration or Vmax is reach at very
high substrate concentrations [S] (at saturation)
 Effect of Substrate Concentration
v
At saturation [S] saturation, all the E are ES

Vmax

Vmax/2

When [S] increase, part of the enzyme become

involved as ES and a part remains free
E= ES + E free (remaining)

0 Km
[S]
At [S]= 0, all the E are free and no reaction
Initial rate or velocity (v) at any time= Kcat.[ES]
So Vmax= Kcat. [Et] since all the enzymes will be active and
catalyzing the reaction
 Michaelis–Menten Equation
Example of Hexokinase and Glucokinase
Lineweaver-Burk Plot
(Double Reciprocal)

y= ax+b

1/V=Km +[S] / Vmax.[S]

 Enzymes inhibitor
Reversible and Irreversible

Reversible inhibition
 Competitive
 Noncompetitive inhibition.
 Uncompetitive
 Enzymes inhibitor
Competitive inhibitors:

E+S ES E+P
+I
The inhibitor compete with substrate
to bind the enzyme
EI

Vmax Remains Constant

Km

Important!!!

E+ S ES E+P
 Enzymes inhibitor
Noncompetitive inhibitors:
E+S ES E+P
+I +I

EI ESI
Lineweaver-Burk plot
1/V

Km Remains Constant
Vmax -1/Km
1/Vmax

1/[S]
 Enzymes inhibitor
Uncompetitive inhibitors:
E+S ES E+P
Binding of the inhibitor and the increase in the ESI
+I complex may also affect the dissociation of
substrate, causing an apparent decrease in Km, i.e.
ESI an apparent increase in substrate affinity.

1/V
Lineweaver-Burk plot

Km
Vmax
1/Vmax
-1/Km

1/[S]
 Irreversible inhibitors
Irreversible inhibitors are inhibitor that binds the enzyme and modify its active
site irreversibly

Example:
Arachidonate Prostaglandin (Inflammatory response)
Cyclooxygenase

Serine acetylation
Aspirin (acetylsalicylic acid)

Alcohol Acetaldehyde Acetic acid

alcohol dehydrogenase aldehyde dehydrogenase

Cysteine modification
Disulfiram
 Enzyme regulation

■ The expression of the enzyme protein.

■ Enzymes may be irreversibly activated or inactivated

by proteolytic enzymes (Proenzyme or Zymogen).

■ Enzymes may be reversibly activated or inactivated by

covalent modification, such as phosphorylation.

■ Allosteric regulation

■ The degradation of enzymes

 Medical Cases
ACE (Angiotensin Converting Enzyme) inhibitors

Angiotensinogen Angiotensin I Angiotensin II

Renin ACE
(Kidney)

ACE inhibitor (Captopril), Increase Blood

competitive inhibitor Pressure

Methanol Poisoning

Symptoms: visual disturbance and severe metabolic acidosis

Methanol Formate (Toxic) Ethanol Acetic Acid

alcohol dehydrogenase alcohol dehydrogenase