Enzymes

Types of Inhibition
• Competitive Inhibition
• Noncompetitive Inhibition
• Uncompetitive Inhibition
• Irreversible Inhibition
 - Inhibitor
Vmax
R eactio n R ate
+ Inhibitor
Vmax
2 Vmax,app = Vmax
Km,app > Km

Km Km,app
[Substrate]

E+S ES E+P
+
I

EI
 Practical example
alcohol dehydrogenase
Methanol formaldehyde formic acid

Methanol (CH3OH) is metabolized to formaldehyde and formic

acid by alcohol dehydrogenase. Since ethanol (CH3CH2OH)
competes with methanol for the same binding site on alcohol
dehydrogenase, it slows the metabolism of methanol,
allowing the toxic metabolites to be disposed of before they
build up to dangerous levels.
 Example of a Competitive Inhibitor
• Malonate is a competitive inhibitor of succinate dehydrogenase
- it has a structure that is similar to succinate
- inhibition can be reversed by adding succinate
Reaction Rate Vmax - Inhibitor

Vmax,app
1
V
+ Inhibitor
2 max
1
V
Vmax,app < Vmax
2 max,app
Km,app = Km

Km [Substrate]
Km,app

E+S ES E+P
+ +
I I

EI + S ESI
 Example of noncompetitive inhibition: fructose 1,6-
bisphosphatase inhibition by AMP

Fructose 1,6-bisphosphatase is a key regulatory enzyme in the

gluconeogenesis pathway. High amounts of AMP signal that
ATP levels are low and gluconeogenesis should be shut down
while glycolysis is turned on.

High AMP levels inhibit fructose 1,6-bisphosphatase (shutting

down gluconeogenesis) and activate phosphofructokinase
(turning on glycolysis). Regulation of fructose 1,6-
bisphosphatase and phosphofructokinase by AMP prevents a
futile cycle in which glucose is simultaneously synthesized and
broken down.
 Vmax - Inhibitor
Reaction Rate

Vmax,app
1
V + Inhibitor
2 max
1
V
2 max,app Vmax,app < Vmax
Km,app < Km

Km,app Km [Substrate]

E+S ES E+P
+
I

ESI
Examples of Irreversible Inhibitors

• diisopropylphosphofluoridate
– permanently inactivates serine proteases by
forming a covalent bond with the active site
serine
 Summary-Enzyme Inhibition
• Competitive Inhibitor
– Binds to substrate binding site
– Competes with substrate
– The affinity of the substrate appears to be decreased
when inhibitor is present (Km,app >Km)
• Noncompetitive inhibitor
– Binds other than substrate binding site
– Does not compete with the substrate for binding to
the enzyme
– The maximum velocity appears to be decreased in the
presence of the inhibitor (Vmax,app <Vmax)
• Uncompetitive Inhibitor
– Binds to the enzyme only after the substrate has
bound
– The affinity of the substrate appears to be increased
and the maximum velocity appears to be decreased
when inhibitor is present (Km,app <Km, Vmax,app <Vmax),
• Irreversible Inhibitor
– Covalently modifies and permanently inactivates the
enzyme
Bi-substrate single displacement reaction

• Consider the case of an enzyme catalyzing a

reaction involving two substrates, A
and B, and yielding the products P and Q:

• Such a reaction is termed a bi-substrate reaction.

 random, where either A or B may bind to the enzyme first, followed by the
other substrate

An example of a random, single-displacement mechanism is seen in the enzyme

creatine kinase, a phosphoryl transfer enzyme that uses ATP as a phosphoryl donor to
form creatine phosphate (CrP) from creatine (Cr). Creatine-P is an important reservoir
of phosphate-bond energy in muscle cells
ordered, where A, designated the leading substrate, must bind to E first before
B can be bound.

(A) (B) (P) (Q)

 Double-Displacement bi-substrate (Ping-Pong) Reaction

Glutamate aspartate aminotransferase, an enzyme conforming to a double-displacement

bi-substrate mechanism
 Isoenzymes

• Isoenzymes are different forms of an enzyme that catalyze

the same reaction in different tissues in the body
- they have slight variations in the amino acid sequences
of the subunits of their quaternary structure
• For example, lactate dehydrogenase (LDH), which converts
lactate to pyruvate, consists of five isoenzymes
 Are All Enzymes Proteins?
• RNA Molecules That Are Catalytic Have Been
Termed “Ribozymes”

They are substrate specific, they enhance the reaction rate, and they emerge from the
reaction unchanged.
Most ribozymes act in RNA processing, cutting the phosphodiester backbone at specific
sites and religating needed segments to form functional RNA strands while discarding
extraneous pieces.

For example, bacterial RNase P is a ribozyme involved in the formation of mature tRNA
molecules from longer RNA transcripts.
Antibody Molecules Can Have Catalytic Activity

• Catalytic antibodies are antibodies with catalytic activity called abzymes

• Like other antibodies, catalytic antibodies are elicited in an organism in response to
immunological challenge by a foreign molecule called an antigen
• In this case, however, the antigen is purposefully engineered to be an analog of the
transition state in a reaction.

• Thus, a catalytic antibody facilitates, or catalyzes, a reaction by forcing the

conformation of its substrate in the direction of its transition state.