Purification and characterization of the cystinyl bond cleaving yeast aminopeptidase yscXVI.

Abstract:

:Aminopeptidase yscXVI was purified from the yeast Saccharomyces cerevisiae. By SDS-PAGE the enzyme has a molecular weight of 45,000 Da, and in chromatofocusing, elution was observed at pH 6.2. The synthetic substrate cystinyl-4-nitroanilide (Km 22.5 microM, Vmax 12.9 mU/mg) is cleaved most efficiently in the pH range 7-8. Besides cleaving this standard substrate, aminopeptidase yscXVI acts on several other 4-nitroanilide substrates with unsubstituted N-terminal L-amino acids. Highest hydrolysis rate was measured with Lys-4-nitroanilide and Leu-4-nitroanilide. The activity of aminopeptidase yscXVI is abolished by chelating agents and restored by Zn2+, Mn2+ and Co2+ ions. Bestatin and amastatin are both strong inhibitors of the enzyme, with Ki values of 0.53 microM and 0.93 microM, respectively. Aminopeptidase yscXVI is detectable in the logarithmic growth phase, stationary phase, and in starved cultures of yeast.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Tisljar U,Wolf DH

doi

10.1016/0014-5793(93)81566-i

subject

Has Abstract

pub_date

1993-05-10 00:00:00

pages

191-6

issue

2

eissn

0014-5793

issn

1873-3468

pii

0014-5793(93)81566-I

journal_volume

322

pub_type

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