Chapter 6
The Behavior of
Proteins: Enzymes
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Enzymes
• Biological catalysts, usually globular proteins
• Increase the rate of a reaction by a factor of up to
1020 over an uncatalyzed reaction
• Highly specific to the extent that they can distinguish
stereoisomers of a given compound
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Thermodynamic Principles - Enzyme Catalysis
• Standard free energy change (ΔG°): Difference
between energies of reactants and products under
standard conditions
• Enzymes can speed up reactions but cannot alter the
equilibrium constant or the free energy change
• Reaction rate depends on its activation energy
(ΔG°‡), which is the energy input required to initiate a
reaction
• ΔG°‡ for an uncatalyzed reaction is higher than that for
a catalyzed reaction
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The Transition State
• Intermediate stage in a reaction in which old bonds
break and new bonds are formed
• In the plot of the energies for a spontaneous reaction,
transition state lies at the maximum of curve
connecting the reactants and products
• ΔG°‡ can be the amount of free energy required to
bring the reactants to the transition state
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The Transition State (continued)
• ΔG° of a reaction remains unchanged when a
catalyst is added, but ΔG°‡ is lowered
• Presence of an enzyme lowers ΔG°‡ needed for
substrate molecules to reach the transition state
• Concentration of the transition state increases
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Temperature Dependence of Catalysis
• Increasing the temperature of a reaction mixture will
increase the energy available to the reactants to
reach the transition state
• Occurs only to a limited extent with biochemical
reactions because increasing temperature will
eventually lead to enzyme denaturation
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Enzyme–Substrate Binding
• In an enzyme-catalyzed reaction:
• Substrate (S) is a reactant
• Active site: Portion of the enzyme surface to which
the substrate binds via noncovalent forces and at
which the reaction takes place
• Noncovalent forces include hydrogen bonding,
electrostatic attractions, and van der Waals attractions
• First step is binding of substrate to the enzyme
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Enzyme–Substrate Binding Models
• Lock-and-key model
• Substrate binds to that portion of the enzyme with a
complementary shape
• Induced fit model
• Binding of the substrate induces a change in the
conformation of the enzyme that results in a
complementary fit
• Binding site has a different three-dimensional shape
before the substrate is bound
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Two Models for the Binding of a Substrate to an
Enzyme
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Formation of Product from Substrate
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Inhibitors
• Substances that decrease the rate of an enzyme-
catalyzed reaction
• Reversible inhibitors
• Substances that bind to an enzyme and subsequently
are released
• Include competitive, noncompetitive, and
uncompetitive inhibitors
• Irreversible inhibitors
• Substances that react with enzymes to produce
proteins that are not enzymatically active and from
which original enzymes cannot be regenerated
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Competitive Inhibition
• Decrease in enzymatic activity caused by binding of a
substrate analogue to the active (catalytic) site
• Inhibitor competes with the substrate for the active
site on the enzyme
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Substrate or Inhibitor Binding in the Case of
Competitive Inhibition
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Identifying a Competitive Inhibitor
• In the presence of a competitive inhibitor:
• Slope of the Lineweaver–Burk plot changes
• y intercept of the graph does not change
• Vmax is unchanged
• KM increases by the following factor:
1+
I
KI
• Substitute value of KM in the following equation:
1 KM 1 1
= × +
V Vmax S Vmax
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Identifying a Competitive Inhibitor (continued)
1 KM 1 1 1
= 1 + × +
V Vmax K I S Vmax
y = m × x + b
• In a Lineweaver–Burk double-reciprocal plot of 1/V
versus 1/[S], the slope and the x intercept change
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Lineweaver–Burk Double-Reciprocal Plot of Enzyme
Kinetics for Competitive Inhibition
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Noncompetitive Inhibition
• Form of enzyme inactivation in which a substance
binds to a site other than the active site but distorts
the active site to inhibit reaction
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Nature of Substrate and Inhibitor Binding in
Noncompetitive Inhibition
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Identifying a Noncompetitive Inhibitor
• Inhibitor does not interfere with substrate binding
• Value of Vmax decreases, and value of KM remains the
same
• Increasing substrate concentration cannot overcome
noncompetitive inhibition
• Inhibitor and substrate are not competing for the same
site
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Lineweaver–Burk Plot of Enzyme Kinetics for
Noncompetitive Inhibition
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Irreversible Inhibition
• Covalent binding of an inhibitor to an enzyme,
causing permanent inactivation
• Suicide substrates: Molecules used to bind to an
enzyme irreversibly and inactivate it
• Used in medicine
• Example - Antibiotic penicillin
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