Last Update: 2 November Part I

2017 M - 25
Inhibition of enzymes action
Inhibition of an enzyme is a decline in the rate of its catalytic activity, resulting from the binding of
sum low molecular weight ligands, called inhibitor. Inhibitor alters the substrate affinity or maximum
velocity of an enzyme. Basically, enzyme inhibition is two types,
(i) reversible inhibition where enzyme activity is restored form enzyme inhibitor complex and
the binding of inhibitor with enzyme is weak and easily dissociable non-covalent bonds.
(ii) irreversible enzyme inhibition is one of the system where enzyme fails to free form enzyme
inhibitor complex. Inhibitor is bound with enzyme by stable covalent bonds.
According to the stability of enzyme inhibitor complex we can classify the enzyme inhibition in to 3
groups ,
(i) competitive,
(ii) non-competitive,
(iii) uncompetitive.

1. Competitive inhibition:
It is also called substrate analogue inhibition. It is reversible inhibition. The inhibitor binds to the
enzyme by weak and easily dissociable bonds. Example, The enzyme succinate dehydrogenase has a
competitive inhibitor malonate

Characteristics of competitive inhibitor.

i) The inhibitor bears a close structural similarity with the substrate and competes for the
substrate binding site of the enzyme.
ii) After binding, inhibitor forms EI (enzyme-inhibitor) complex that lowers the substrate
affinity of that enzyme.
iii) The lowering of substrate affinity by the inhibitor in reflected in the rise of in K m value.
The altered Km value is called apparent Km or Km
iv) A higher concentration of the substrate may overcome competitive inhibition. So high
substrate concentration gives same Vmax. For this reason in competitive inhibition Vmax is
unchanged
v) The magnitude of competitive inhibition depends on the relative concentration of the
substrate and inhibitor.

Kinetics:
Competitive inhibition lowers Vo,
[I]
increases Km to Km 1 and does
Ki
not affect Vmax.
Where, [I] = Molar concentration of
inhibitor, Ki = dissociation constant of
the EI complex.

a) The Michaelis Menten equation

is modified like that by competitive

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inhibition.
Vmax [S]
Vo =
[I]
K m 1 [S]
Ki

b) Equation for double reciprocal plot

1 K I 1 1
m 1 .
Vo Vmax K i [s] Vmax

1
Y intercept =
Vmax

1
X intercept =
[I]
K m 1
Ki

Km I
Slop = 1
Vmax K i

c) Equation for Eadie Hofstee plot

Vo [I]
Vo Vmax . K m 1
[S] Ki

Y intercept = Vmax

Vmax
X intercept =
[I]
K m 1
Ki

[I]
Slop = -Km 1
Ki

d) Equation for Wolf-Hanes plot

[S] K m [I] 1
1 +[S]
Vo Vmax Ki Vmax

Km [I]
Y intercept = 1
Vmax Ki

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 [I]
X intercept = -Km 1
Ki
1
Slop
Vmax
2. Noncompetitive inhibition:
Non Competitive inhibition may be either reversible or irreversible. It depends on the character of
the inhibitor. The major characteristic of non competitive inhibition-
(i) The inhibitors has no structural similarity with substrate.
(ii) It binds with enzyme at the site other than substrate binding site and probably it changes
the 3 dimensional conformation of the enzyme, resulting in a failure to release the
products from enzyme substrate complex.
(iii) Though inhibitor does not compete with substrate binding site. Therefore Km unchanged.
(iv) Inhibitors may bind with free enzyme or ES complex to form EI or ESI complex
respectively.
(v) The inhibitor cannot dislodged from enzyme inhibitor complex by increasing the
substrate concentration. So Vmax cannot reach its original value. Thus in this inhibition
[I]
Vmax is reduced by a factor 1
Ki
Vmax
Thus the altered Vmax reduced to
[I]
1
K i
depending on inhibitor concentration [I].

Changes in kinetics
Michaelis Menten equation is modified like that

Vmax [S]
Vo =
[I]
(K m [S]) 1
Ki

The double reciprocal plot equation is

1 K [I] 1 1 [I]
m 1 . 1
Vo Vmax K i [s] Vmax K i

1 [I]
Y intercept = 1
Vmax K i

1
X intercept =
Km
Km [I]
Slop = 1
Vmax Ki

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ii In Eadie Hofstee plot the equation is

Vmax V
Vo = o Km
[I] [S]
1
Ki

Vmax
Y intercept =
[I]
1
Ki

Vmax
X intercept =
[I]
K m 1
Ki
Slop = - Km

iii The Wolf Hanes plot equation is

[I]
1
[S] K m [I] Ki
1 [S]
Vo Vmax Ki Vmax

Km [I]
Y intercept = 1
Vmax Ki

X intercept = -Km

1 [I]
Ki
Slop =
Vmax

3 Uncompetitive inhibition :
Uncompetitive inhibition is reversible
inhibition occurs in many by substrate or multi
substrate reactions. The major characteristic of
these inhibitions are as follows:
(i) Inhibitor has no structural similarity of the
substrate
(ii) The inhibitors neither bind to free enzyme.
It binds other than active site. Inhibitors
can only bind to ES complex and form
ESI complex. There by it enhances the
substrate affinity of the enzyme by
changing enzyme conformation and
inhibits the release of product from the
enzyme and lowers both Km and Vmax.

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 Vmax Km
(iii) Vmax is altered to and Km changes into
[I] [I]
1 1
Ki Ki
Changes in kinetics.
In uncompetitive inhibition the Michaelis Menten
equation changes to
Vmax [S]
Vo =
[I]
Km [S] 1
Ki

i. In double reciprocal plot:

1 K 1 1 [I]
m 1
Vo Vmax [S] Vmax K i

1 [I]
Y intercept = 1
Vmax K i

1 [I] Km
Slop =
Vmax
X intercept = - K i
Km

ii Eadie Hofstee plot

Vmax Vo Km
Vo = .
[I] [S] [I]
1 1
Ki Ki

Vmax
Y intercept =
[I]
1
Ki

Vmax Km
X intercept = Slop =
Km [I]
1
Ki
iii Wolf Hanes plot:

[S] K m 1 [I]
[S] 1
Vo Vmax Vmax Ki

Km Km 1 [I]
Y intercept = X intercept =
Vmax [I]
1 Slop = Ki
Ki Vmax

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