生物化学课件§5 Enzyme.pptVIP

liguofu Many enzymes need non-protein cofactors to help in catalysis For E + S ES E + P Km = If k 2 is rate-limiting, k 2k -1, Km = k -1 /k 1 Km equals to the dissociation constant (Kd) of the ES complex; Km represent a measure of affinity of the enzyme for its substrate in the ES complex. If k 2 k -1, then Km = k 2 / k1 If k2 and k-1 are comparable, Km is a complex function of all three rate constants. The value of kcat/Km has an upper limit (for the perfected enzymes) It can be no greater than k1 The decomposition of ES to E + P can occur no more frequently than that E and S come together to form ES. The most efficient enzymes have kcat/Km values near the diffusion-controlled limit of 108 to 109 M-1S-1. Each substrate will have one characteristic Km value. Ternary complex may or may not be formed for the bisubstrate reactions depending on the mechanism. Steady-state kinetics can often help distinguish these two mechanisms. A second catalytically important residue, His57, was discovered by an affinity iactivator (TPCK) Chymotrypsin (and other serine proteases) acts via a mixture of covalent and general acid-base catalysis to cleave (not a direct attack of water on the peptide bond!) The specificity of serine proteases is determined by the structural features of a substrate binding pocket The activities of enzymes can be regulated via mainly three principle ways Allosteric regulation (noncovalent modifications, reversible); Covalent modifications (reversible); Proteolytic cleavage (irreversible). (Gene regulation: changing the amount of specific enzymes). Allosteric modulators can be either inhibitory or stimulatory Allosteric enzymes are often oligomeric: ATCase consists of two catalytic trimers and three regulatory dimers Multiple phosphorylations can occur on one enzyme: allowing subtle modulation of activity. Some regulatory enzymes use multiple regulatory mechanisms The bacterial Gln synthetase is r