|
type |
Journal Article |
authors |
Ono, B.; Ishii, N.; Naito, K.; Miyoshi, S.; Shinoda, S.; Yamamoto, S.; Ohmori, S. |
title |
Cystathionine gamma-lyase of Saccharomyces cerevisiae: structural gene and cystathionine gamma-synthase activity |
journal |
Yeast |
Activity |
4.4.1.1 |
Family |
4.4.1.1 |
sel |
selected |
ui |
8511969 |
year |
(1993) |
volume |
9 |
number |
4 |
pages |
389-97 |
| |
keywords |
Amino Acid Sequence |
abstract |
Purification of Saccharomyces cerevisiae cystathionine gamma-lyase (gamma-CTLase) was hampered by the presence of a protein migrating very close to it in various types of column chromatography. The enzyme and the contaminant were nevertheless separated by polyacrylamide gel electrophoresis. N-terminal amino acid sequence analysis indicated that they are coded for by CYS3 (CYI1) and MET17 (MET25), respectively, leading to the conclusion that CYS3 is the structural gene for gamma- CTLase and that the contaminant is O-acetylserine/O-acetylhomoserine sulfhydrylase (OAS/OAH SHLase). Based on these findings, we purified gamma-CTLase by the following strategy: (1) extraction of OAS/OAH SHLase from a CYS3-disrupted strain; (2) preparation of antiserum against it; (3) identification of a strain devoid of the OAS/OAH SHLase protein using this antiserum; and (4) extraction of gamma-CTLase from this strain. Purified gamma-CTLase had cystathionine gamma-synthase (gamma-CTSase) activity if O-succinylhomoserine, but not O- acetylhomoserine, was used as substrate. From this notion we discuss the evolutional relationship between S. cerevisiae gamma-CTLase and Escherichia coli gamma-CTSase. |
last changed |
2009/06/30 19:50 |
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