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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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