Isolation,and biochemical and biological characterization of an a-mating-type-specific glycoprotein responsible for sexual agglutination from the cytoplasm of a-cells,in the yeast Saccharomyces cerevisiae |
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Authors: | Yamaguchi Makoto Yoshida Kazuo Yanagishima Naohiko |
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Institution: | (1) Biological Institute, Faculty of Science, Nagoya University, Chikusa-ku, 464 Nagoya, Japan;(2) Present address: Central Institute for Medical Research, Nissui Pharmaceutical Co., Ltd., 1075-2, Hokunanmoro, Yuukishi, Ibaraki, Japan |
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Abstract: | An a-mating-type-specific substance responsible for sexual agglutination was purified to 397-times in specific activity (units/mg protein) from the cytoplasm of a-mating type cells. The purified substance gave a single band stained with PAS reagent but not with both Coomassie brilliant blue and silver staining reagent by polyacrylamide gel electrophoresis in the presence of 8 M urea. However, incorporation of 35S]methionine and Lowry reaction clearly indicate that the substance is a glycoprotein. The substance specifically masked sexual agglutinability of cells of the opposite mating type , indicating univalent action. The substance is a glycoprotein with a carbohydrate content of 90%, a pI of 4.5, and a molecular weight of 130,000. The substance was inactivated by 2-mercaptoethanol and proteolytic enzymes but not by glycolytic enzymes. The substance formed a complementary complex having no biological activity when mixed with -agglutination substance from the wall or cytoplasm of -cells in vitro.Non-common abbreviations PAGE
polyacrylamide gel electrophoresis
- PAS
periodic acid-Schiff
- PBS
10-2 M phosphate buffer solution, pH 5.5
- PMSF
phenylmethyl sulfonyl fluoride
- SDS
sodium dodecyl sulfate |
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Keywords: | Agglutination substance Cell-cell recognition Glycoprotein Mating Saccharomyces cerevisiae Sexual agglutinability Yeast |
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