SPECIFICITY OF

ENZYMES
• In an enzyme-catalyzed reaction,
the enzyme binds to the substrate
(one of the reactants) to form a
complex. The formation of the
complex leads to the formation of
the transition-state species, which
then forms the product. A substrate
binds, usually by non-covalent
interactions, to a small portion of
the enzyme called the active site,
frequently situated in a cleft or
crevice in the protein and
consisting of certain amino acids
that are essential for enzymatic
activity.
Active Site
• is a specific region on the enzyme that binds to the substrate. The active
site is a 3-dimensional crack in the enzyme that contains the catalytic
groups – the residues part of the enzyme that are responsible for catalyzing
the reaction.
• contains catalytic residues responsible for lowering the free energy of the
transition state and stimulate bond breaking/forming.
Active Site cont’d..
• establishes micro-environments. Active sites create non-polar
environments unless water participates in the reaction. This also prevents
unwanted reactions.
• only make up of a very small portion of the enzyme.
• typically bind substrates reversibly via non-covalent forces.
• has structures complementary to their corresponding substrates.
Lock-and-Key Model
• first postulated in 1894 by Emil Fischer.
• the lock is the enzyme and the key is the substrate.
• assumes a high degree of similarity between the shape of the
substrate and the geometry of the binding site on the enzyme.
• the substrate binds to a site whose shape complements its own,
like a key in a lock or the correct piece in a three-dimensional
jigsaw puzzle.
• thismodel has intuitive appeal but is now largely of historical
interest because it does not take into account an important
property of proteins, namely their conformational flexibility.
Induced-Fit Model
• proposed by Daniel Koshland in 1958.
• the binding of the substrate induces a conformational change in
the enzyme that results in a complementary fit after the
substrate is bound.
• The binding site has a different three-dimensional shape before
the substrate is bound.
• It suggests that the active site continues to change until
the substrate is completely bound to it, at which point the final
shape and charge is determined.
Induced-Fit Model cont’d..

• more accepted model for enzyme-substrate complex than the

lock-and-key model.
• shows that enzymes are rather flexible structures in which the
active site continually reshapes by its interactions with
the substrate until the time the substrate is completely bound to
it.
What would happen if this binding were too perfect? The figure shows what happens when E
and S bind. An attraction must exist between E and S for them to bind. This attraction causes the
ES complex to be lower on an energy diagram than the E + S at the start. Then the bound ES must
attain the conformation of the transition state EX‡ . If the binding of E and S to form ES were a
perfect fi t, the ES would be at such a low energy that the difference between ES and EX‡ would
be very large. This would slow down the rate of reaction. Many studies have shown that enzymes
increase the rate of reaction by lowering the energy of the transition state, EX‡ , while raising the
energy of the ES complex. The induced-fit model certainly supports this last consideration better
than the lock-and-key model; in fact, the induced-fit model mimics the transition state.
Lock-and-Key VS. Induced-Fit Model
Some Digestive Enzymes and their Substrates
Enzyme Substrate
Deoxyribonuclease DNA
Dipeptidase Dipeptides
Lactase Lactose
Sucrase Sucrose
Ribonuclease RNA
Maltase Maltose
Amylase Amylose