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CHEM464/Medh, J.D.
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CHEM464/Medh, J.D.
Significance of MM equation
• KM has a unit of concentration. It is a constant for every Line-Weaver Burk Plot
enzyme under specific conditions
• Also called the double-reciprocal plot
• When [S] <<< KM, Vo is directly proportional to [S]
• When [S] >>> KM, Vo = Vmax
• 1/Vo = (KM/Vmax).(1/[S]) + 1/Vmax
• When [S] = KM, Vo = ½ Vmax • This is equation for a straight line y = mx + c
• The KM value of an enzyme indicates the concentration of the • Slope = KM/Vmax
substrate required for significant catalysis
• Since KM = k2+k-1/k1, when k-1 >>> k2, KM = k-1/k1 • Y-intercept = 1/Vmax
• Dissociation constant of ES, KES = [E][S] / [ES] = k-1/k1 • X-intercept = -1/KM
• When k-1 >>> k2, KM = KES. High KM = high dissociation =
• Useful for experimental determination of KM and Vmax
weak binding between E and S
• When [E]T = [ES], Vmax = k2[E]T. k2 is called the turnover
number: the rate when enzyme is saturated with substrate
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CHEM464/Medh, J.D.
Irreversible Inhibitors
Uncompetitive Inhibitors
• Group-specific: covalently modify specific side
• The inhibitor cannot bind to the enzyme directly, chains of aa in active site of enzyme. Eg: DIPF
but can only bind to the enzyme-substrate reacts with serine, iodoacetamide modifies cys
complex. • Some substrates transiently covalently modify the
• ES + I = ESI. enzyme. Substrate analogs are structurally similar
to such substrates. Since they are not actually
• Both Vmax and KM are decreased by (1+I/Ki). substrates, the enzymatic reaction cannot proceed.
• Suicide inhibitors are modified substrates that trick
the enzyme into initiating the reaction. Instead of
product, reactive intermediates are released which
react with the enzyme causing its inactivation
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CHEM464/Medh, J.D.