General occurrence of binding synergism in zinc proteases and its possible significance.
The observation of binding synergism has been successfully extended to include carboxypeptidases A and B. The behaviour of these two enzymes follows the same pattern previously found for three other Zn-proteases. Thus in all cases examined, the affinity of a suitable Zn-ligand is increased in the presence of a compound (specificity probe) which contains the key structural features of specific substrates. A bifunctional ligand such as phosphonoacetate is particularly useful for generating synergism in both carboxypeptidases. Presumably the carboxylate moiety binds to the C-terminal recognition site while the other functional group interacts with the metal ion. Several basic compounds (e.g. methyl guanidine) act as effective specificity probes for carboxypeptidase B while phenol and other hydrophobic substances serve this purpose in carboxypeptidase A. The above phenomenon appears to be a mechanism designed to enhance catalytic efficiency through a substrate-induced conformational change. We postulate that there is a requirement for at least one ionizable group at the active site. The proposed mechanism keeps this group in the correct ionization state in the presence of water and increases its reactivity after exclusion of water by substrate binding. We suggest the term xerophilic shift for this process. Since proton transfer is a common process in enzyme reactions, the xerophilic-shift mechanism may play a similar role in many instances. It should therefore be possible to detect binding synergism in a wide variety of enzymes.
Chan
W W
WW
Department of Biochemistry, McMaster University, Hamilton, Canada.
Pfuetzner
R A
RA
D000818:Animals; D001665:Binding Sites; D043424:Carboxypeptidase B; D002268:Carboxypeptidases / Q000378:metabolism*; D043422:Carboxypeptidases A; D002417:Cattle; D003432:Cross-Linking Reagents; D008024:Ligands; D010179:Pancreas / Q000201:enzymology; D013552:Swine; D015032:Zinc / Q000378:metabolism*
D003432:Cross-Linking Reagents; D008024:Ligands; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D043424:Carboxypeptidase B; D015032:Zinc