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Aminopeptidase yscCo‐II: a new cobalt‐dependent aminopeptidase from yeast—purification and biochemical characterization
Author(s) -
HerreraCamacho Irma,
MoralesMonterrosas Rosalva,
QuirózAlvarez Rubén
Publication year - 2000
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(200002)16:3<219::aid-yea523>3.0.co;2-j
Subject(s) - aminopeptidase , cobalt , enzyme , size exclusion chromatography , biology , pmsf , biochemistry , yeast , enzyme assay , chromatography , molecular mass , leucyl aminopeptidase , chemistry , amino acid , leucine , organic chemistry
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. Copyright © 2000 John Wiley & Sons, Ltd.