Chloride as allosteric effector of yeast aminopeptidase I.
Activation of yeast aminopeptidase I by chloride was studied by kinetic methods. Several effects contributed to overall activity enhancement: At low concentrations of Zn2+ (an essential component of aminopeptidase I) chloride increased the amounts of active enzyme by reducing the cooperativity of metal binding. In addition, substrate turnover was enhanced due to increased kcat and a moderate decrease of Km. At high concentrations of Zn2+ substrate saturation curves were sigmoidal. Under these conditions chloride activated by restoring Michaelis-Menten kinetics of substrate turnover. At the same time, reconstitution of active enzyme from apoprotein and Zn2+ was substantially accelerated and its inactivation due to loss of Zn2+ was retarded. Co2+-Substituted aminopeptidase I, although catalytically active, was much less sensitive to chloride activation. Apparent binding constants for chloride, as estimated from its effects on metal binding and catalysis, respectively, were different. This suggests that two independent activation mechanisms may be operative. Both appear to be mediated by conformational changes of the enzyme protein.
D000494:Allosteric Regulation; D000626:Aminopeptidases / Q000378:metabolism*; D002712:Chlorides / Q000494:pharmacology*; D004789:Enzyme Activation / Q000187:drug effects; D006868:Hydrolysis; D007700:Kinetics; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins; D013379:Substrate Specificity; D015032:Zinc / Q000378:metabolism
D002712:Chlorides; D029701:Saccharomyces cerevisiae Proteins; D000626:Aminopeptidases; C019689:APE1 protein, S cerevisiae; D015032:Zinc