TY - JOUR
T1 - Characterization of KEX2-encoded endopeptidase from yeast Saccharomyces, cerevisiae
AU - Mizuno, Kensaku
AU - Nakamura, Tomoko
AU - Ohshima, Takehiro
AU - Tanaka, Shoji
AU - Matsuo, Hisayuki
PY - 1989/2/28
Y1 - 1989/2/28
N2 - Yeast Saccharomyces, cerevisiae, KEX2 gene previously isolated was characterized as the gene encoding an endopeptidase required for proteolytic processing of precursors of α-factor and killer toxin. In this study, the cloned KEX2 gene was introduced into the kex2 mutant cells and the KEX2 gene product expressed in these cells was partially purified from their membrane fraction. The enzyme preparation exhibits a calcium-dependent endopeptidase activity with a substrate specificity toward the carboxyl side of Lys-Arg, Arg-Arg and Pro-Arg sequences. The enzyme activity was inhibited by serine-protease inhibitors, such as DFP and PMSF, indicating that the KEX2 endopeptidase belongs to a serine-protease family. The optimal pH was determined to be around 5.5. Thus, the KEX2 endopeptidase was found to be a unique calcium-dependent serine-protease distinct from calpain and trypsin.
AB - Yeast Saccharomyces, cerevisiae, KEX2 gene previously isolated was characterized as the gene encoding an endopeptidase required for proteolytic processing of precursors of α-factor and killer toxin. In this study, the cloned KEX2 gene was introduced into the kex2 mutant cells and the KEX2 gene product expressed in these cells was partially purified from their membrane fraction. The enzyme preparation exhibits a calcium-dependent endopeptidase activity with a substrate specificity toward the carboxyl side of Lys-Arg, Arg-Arg and Pro-Arg sequences. The enzyme activity was inhibited by serine-protease inhibitors, such as DFP and PMSF, indicating that the KEX2 endopeptidase belongs to a serine-protease family. The optimal pH was determined to be around 5.5. Thus, the KEX2 endopeptidase was found to be a unique calcium-dependent serine-protease distinct from calpain and trypsin.
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U2 - 10.1016/0006-291X(89)92438-8
DO - 10.1016/0006-291X(89)92438-8
M3 - Article
C2 - 2647083
AN - SCOPUS:0024513075
VL - 159
SP - 305
EP - 311
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 1
ER -