The effect of monovalent cations on the pre-steady state reaction kinetics of bovine activated plasma protein C and des-1-41-light chain activated plasma protein C.
A pre-steady state kinetic analysis of the stimulation by monovalent cations of the activity of bovine activated protein C (APC) and a proteolytic fragment of APC, des-1-41-light chain activated protein C (GDAPC), toward the substrate, 4-methylumbelliferyl p-guanidinobenzoate, has been undertaken. With the cations Na+ and Cs+, at least two cation sites, or classes of sites, on APC were found to be important to the kinetic effects observed. For GDAPC, with both monovalent cations investigated, a single cation-binding site, or class of sites, of kinetic importance was discovered. The most general mechanism that fits all kinetic data was a rapid equilibrium type, with the cation(s) (A) and substrate (S) binding to the enzyme in a random fashion. Cations were found to be essential activators, and only formation of the EAS or EA2S complex led to product generation. For each enzyme, stimulation of the reaction rates was found to be chiefly due to a dramatic enhancement by monovalent cations of the rate constant (k2) for acylation of the enzyme since the dissociation constant (Ks) for enzyme-substrate interactions was increased in the presence of cations, and the deacylation rate constant (k3) was not affected by these activators.
D000818:Animals; D001665:Binding Sites; D002414:Cations, Monovalent; D002417:Cattle; D002586:Cesium / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D007700:Kinetics; D010446:Peptide Fragments / Q000097:blood*; D011486:Protein C / Q000097:blood*; D012964:Sodium / Q000494:pharmacology*
C017532:4-methylumbelliferylguanidinobenzoate; C050919:7-hydroxy-3-methylcoumarin; D002414:Cations, Monovalent; D010446:Peptide Fragments; D011486:Protein C; D002586:Cesium; D006923:Hymecromone; C050706:protein C, des-1-41 light chain activated-; D012964:Sodium