ACTIVE SITE OF ENZYMES

What are active sites?

• Regions that binds the substrates (cofactor as
the case may be).
• They contain catalytic groups, basically amino
acids that promote formation and degradation
of bonds.
• It enhances the enzyme to bind to substrate
and promote catalysis
 Structure
• The active site has a shape of a three-dimensional
cleft containing amino acids from different residues
of the primary amino acid sequence.

• This active site region is relatively small compared to

the rest of the enzyme.

• The unique amino acids contained in an active site

promote specific interactions that are necessary for
proper binding and resulting catalysis.
• Enzyme specificity depends on the arrangement of
atoms in the active site

• Complementary shapes between enzyme and

substrate(s) allow a greater amount of weak non-
covalent interactions including electrostatic forces,
Van der Waals forces, hydrogen bonding, and
hydrophobic interactions.
 Properties of active sites

• The active site is a three-dimensional cleft formed by

groups that come from different parts of the amino
acid sequence (contact, catalytic, structural, non-
essential).

• The active site takes up a relatively small part of the

total enzyme mass.

• Active sites are clefts or crevices (where water is

usually excluded) to create a microenvironment.
• Substrates are bound to enzymes by multiple weak
attractions such as hydrogen bond, van der waal
forces hydrophobic interactions and electrostatic
interaction (for charged molecules).

• Active sites have complementary structure for their

substrate. The specificity of binding depends on the
precisely defined arrangement of atoms in an active
site.

• Active site help stabilize transition state by lowering

activation energy.
 Models for enzyme-substrate binding

• The lock-and-key model,

• The induced fit model, and

• Transition-state model.
 Lock-and –key model
• Proposed by fisher in 1890.
• This model presumes that the active site of enzyme is
good fit for substrate and does not require change of
structure of enzyme after enzyme binds substrate.
 Induced-fit model
• The involves the changing of the conformation of the active
site to fit the substrate after binding.
• Induced fit is the model such that structure of active site of
enzyme can be easily changed after binding of enzyme and
substrate.
 Transition state model
• This model starts with an enzyme that binds to a
substrate.

• Requires energy to change the shape of substrate.

• Once the shape is changed, the substrate is unbound

to the enzyme, which ultimately changes the shape of
the enzyme.

• An important aspect of this model is that it increases

the amount of free energy.
 Properties that Affect Binding
• • Complementarity

• • Flexibility

• • Surfaces

• • Non-Covalent Forces

• • Affinity