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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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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 α, 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 α-agglutination substance from the wall or cytoplasm of α-cells in vitro.

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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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Author notes

  1. Makoto Yamaguchi

    Present address: Central Institute for Medical Research, Nissui Pharmaceutical Co., Ltd., 1075-2, Hokunanmoro, Yuukishi, Ibaraki, Japan

Authors and Affiliations

  1. Biological Institute, Faculty of Science, Nagoya University, Chikusa-ku, 464, Nagoya, Japan

    Makoto Yamaguchi, Kazuo Yoshida & Naohiko Yanagishima

Authors
  1. Makoto Yamaguchi
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  2. Kazuo Yoshida
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  3. Naohiko Yanagishima
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Yamaguchi, M., Yoshida, K. & Yanagishima, N. 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 . Arch. Microbiol. 140, 113–119 (1984). https://doi.org/10.1007/BF00454912

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  • DOI: https://doi.org/10.1007/BF00454912

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