No abstract

Saccharomyces cerevisiae aminopeptidase yscCo-II (APCo-II) was purified to apparent homogeneity by gel filtration, affinity chromatography and anion-exchange chromatography. APCo-II is an hexameric cobalt-dependent metallo-enzyme with an estimated native molecular mass of 290 kDa. Enzyme activity is only detected in the presence of cobalt ions at pH 7.0. Substrate specificity studies indicate that aminopeptidase yscCo-II cleaves only basic N-terminal residues. PMSF, Cu(2+), 1,10-phenanthroline and bestatin were found to be very strong inhibitors of aminopeptidase yscCo-II activity. Kinetic studies indicated that the enzyme has a similar K(m) and Ka(Co )(activation constant of cobalt) and, following extraction of cobalt from the enzyme, activity was recovered only after cobalt addition.