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It

is a substance which binds with the

enzyme and brings about decrease in the
catalytic activity of the enzyme.

Enzyme inhibitors are molecules that
interact in some way with the enzyme to
prevent it from working in the normal
manner.

Organic or inorganic

Reversible or irreversible
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Reversible

Competitive
Non- competitive
uncompetitive

Irreversible

Allosteric

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Enzyme Inhibition (Mechanism)

Equation and Description

Cartoon Guide

Competitive

Non-competitive

Uncompetitive
E

Substrate

S
I

I
Inhibitor

Compete for
active site

E + S
ES E + P
+
I

EII
[II] binds to free [E] only,
and competes with [S];
increasing [S] overcomes
Inhibition by [II].

Different site
E + S
ES E + P
+
+
I
I

EII + S EIIS
[II] binds to free [E] or [ES]
complex; Increasing [S] can
not overcome [II] inhibition.

E + S
ES E + P
+
I

EIIS
[II] binds to [ES] complex
only, increasing [S] favors
the inhibition by [II].
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Enzyme Inhibition (Plots)

I

Competitive

Non-competitive

Vmax

Double Reciprocal

[S], mM

Km Km

Uncompetitive

Vmax

vo

Direct Plots

vo

Vmax

[S], mM

Km = Km

Vmax

Km Km

Vmax

[S], mM

Vmax unchanged
Km increased

Vmax decreased
Km unchanged

Both Vmax & Km decreased

1/vo

1/vo

1/vo

Intersect
at Y axis

1/Km

Two parallel
lines

1/ Vmax
1/[S]

1/ Vmax

Intersect
at X axis

1/Km

1/[S]

1/ Vmax
1/Km

1/[S]
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Reaction of the irreversible inhibitor

diisopropylfluorophosphate (DFP) with a
serine protease

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Iodoacetate is an irreversible inhibitor of all cysteine

peptidases

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Allosteric protein:-two or more

topological distinct binding
sites which interact functionally
with each other.
Cooperatability:-modification of
the binding constant of the
protein for a small molecule by
the prior binding of another
small molecule.
+ve:-binding

ability or affinity
increases
-ve:- binding ability or affinity
decreases

Allosteric effectors(inhibitors &

activators):- for speed up & to
speed down

when 2,3-BPG binds to an allosteric site

on hemoglobin, the affinity for oxygen of
all subunits decreases.

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Where n is cooperativity and n>1

,Indicates positive cooperativity.

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High substrate concentrations

may
cause inhibition in some
enzymatic
reactions, known as substrate
inhibition

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Assumption

:-A second substrate molecule

can bind to the enzyme when S binds the ES
complex, an unreactive intermediate results.

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Unit of enzyme activity:

Used to measure total units of activity in a given
volume of solution.

Specific activity:
Used to follow the increasing purity of an enzyme
through several procedural steps.

Molecular activity:
Used to compare activities of different enzymes.
Also called the turn-over number (TON = kcat)

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Unit of enzyme activity:

mol substrate transformed/min = unit

Specific activity:
mol substrate/min-mg E = unit/mg E

Molecular activity:
mol substrate/min- mol E = units/mol E

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Temperature

pH

Concentration

Activators

Product

concentration

Time

Radiations

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Reaction rate / arbitrary units

Collision rate of
enzymes and
substrates
Number of
enzymes
remaining
undenatured
o

Temperature / C

Reaction rate / arbitrary units

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Increasing kinetic
energy increases
successful collision
rate

Temperature / C
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Reaction rate / arbitrary units

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Permanent disruption
of tertiary structure
leads to loss of active
site shape, loss of
binding efficiency and
activity

Temperature / C
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Reaction rate / arbitrary units

Optimum temperature

Temperature / C
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The

precise shape of an enzyme (and

hence its active site) depends on the
tertiary structure of the protein

Tertiary structure is held together by
weak bonds (including hydrogen bonds)
between R-groups (or side-chains)

Changing pH can cause these side chains
to ionise resulting in the loss of Hbonding
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Temperature: enzymes work best at an optimum

temperature.
Below this, an increase in temperature provides
more kinetic energy to the molecules involved.
The numbers of collisions between enzyme and
substrate will increase so the rate will too.
Above the optimum temperature, and the
enzymes are denatured. Bonds holding the
structure together will be broken and the active
site loses its shape and will no longer work

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Even if temperature lowered enzyme

cant regain its correct shape

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Reaction rate / arbitrary units

Optimum pH

Either side of the

optimum
pH,
the
gradual ionising of the
side-chains (R-groups)
results in loss of Hbonding, 3D structure,
active site shape loss of
binding efficiency and
eventually
enzyme
activity
pH
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Reaction rate / arbitrary units

Optimum pH

This loss of activity is

only truly denaturation
at extreme pH since
between optimum and
these extremes, the loss
of activity is reversible

pH
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pH:

as with temperature, enzymes have

an optimum pH. If the pH changes much
from the optimum, the chemical nature
of the amino acids can change.
This may result in a change in the bonds
and so the tertiary structure may break
down. The active site will be disrupted
and the enzyme will be denatured.
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