Purification and characterization of aminopeptidase yspI from Schizosaccharomyces pombe.
Aminopeptidase yspI was purified to apparent homogeneity from the fission yeast Schizosaccharomyces pombe. The molecular mass of the native enzyme was estimated to be 184 kDa by gel filtration chromatography. A value of 92 kDa was calculated after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The enzyme is thus a dimer with two identical subunits. Optimum pH for cleavage of synthetic aminoacyl-4-nitroanilides is 7.0. Mercury ions, EDTA and chloroquine were found to be potent inhibitors of aminopeptidase yspI activity. Substrate specificity studies indicate that the purified enzyme cleaves L-lysine-4-nitroanilide with high efficiency.
Arbesú
M J
MJ
Departamento de Biologia Funcional, Facultad de Medicina, Universidad de Oviedo, Spain.
Valle
E
E
Suárez-Rendueles
P
P
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D002413:Cations, Divalent / Q000494:pharmacology; D007700:Kinetics; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012568:Schizosaccharomyces / Q000201:enzymology*; D013379:Substrate Specificity
D002413:Cations, Divalent; D000626:Aminopeptidases; C082560:aminopeptidase yspI