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. 1987 Feb;169(2):475–482. doi: 10.1128/jb.169.2.475-482.1987

Identification of glycoprotein components of alpha-agglutinin, a cell adhesion protein from Saccharomyces cerevisiae.

K Terrance, P Heller, Y S Wu, P N Lipke
PMCID: PMC211801  PMID: 3542959

Abstract

Several glycoproteins which inhibit the agglutinability of Saccharomyces cerevisiae mating type a cells were partially purified from extracts of mating type alpha cells. These proteins, called alpha-agglutinin, were labeled with 125I-Bolton-Hunter reagent. The labeled alpha-agglutinin showed specific binding to a cells. Such specific binding approached saturation with respect to agglutinin or cells and was inhibited in the presence of excess unlabeled alpha-agglutinin. Nonspecific binding was similar in a and alpha cells, was neither saturable nor competable, and was three- to fourfold less than the specific binding to a cells at maximum tested agglutinin concentrations. The major a-specific binding species had a low electrophoretic mobility in sodium dodecyl sulfate-polyacrylamide gels and had an apparent molecular weight of 155,000 by rate zonal centrifugation. Endo-N-acetylglucosaminidase H digestion of the purified glycoprotein complex converted the low-mobility material to four major and several minor bands which were resolved by polyacrylamide gel electrophoresis. All but two minor peptides bound specifically to a cells. Analyses of agglutinin from mnn mutants confirmed the deglycosylation results in suggesting that the N-linked carbohydrate portion of alpha-agglutinin was not necessary for activity.

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Selected References

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  1. Betz R., MacKay V. L., Duntze W. a-Factor from Saccharomyces cerevisiae: partial characterization of a mating hormone produced by cells of mating type a. J Bacteriol. 1977 Nov;132(2):462–472. doi: 10.1128/jb.132.2.462-472.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Burke D., Mendonça-Previato L., Ballou C. E. Cell-cell recognition in yeast: purification of Hansenula wingei 21-cell sexual agglutination factor and comparison of the factors from three genera. Proc Natl Acad Sci U S A. 1980 Jan;77(1):318–322. doi: 10.1073/pnas.77.1.318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Crandall M. A., Brock T. D. Molecular basis of mating in the yeast hansenula wingei. Bacteriol Rev. 1968 Sep;32(3):139–163. doi: 10.1128/br.32.3.139-163.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Duntze W., MacKay V., Manney T. R. Saccharomyces cerevisiae: a diffusible sex factor. Science. 1970 Jun 19;168(3938):1472–1473. doi: 10.1126/science.168.3938.1472. [DOI] [PubMed] [Google Scholar]
  5. Duntze W., Stötzler D., Bücking-Throm E., Kalbitzer S. Purification and partial characterization of -factor, a mating-type specific inhibitor of cell reproduction from Saccharomyces cerevisiae. Eur J Biochem. 1973 Jun;35(2):357–365. doi: 10.1111/j.1432-1033.1973.tb02847.x. [DOI] [PubMed] [Google Scholar]
  6. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  7. Fehrenbacher G., Perry K., Thorner J. Cell-cell recognition in Saccharomyces cerevisiae: regulation of mating-specific adhesion. J Bacteriol. 1978 Jun;134(3):893–901. doi: 10.1128/jb.134.3.893-901.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hagiya M., Yoshida K., Yanagishima N. The release of sex-specific substances responsible for sexual agglutination from haploid cells of Saccharomyces cerevisiae. Exp Cell Res. 1977 Feb;104(2):263–272. doi: 10.1016/0014-4827(77)90090-8. [DOI] [PubMed] [Google Scholar]
  9. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Lipke P. N., Terrance K., Wu Y. S. Interaction of alpha-agglutinin with Saccharomyces cerevisiae a cells. J Bacteriol. 1987 Feb;169(2):483–488. doi: 10.1128/jb.169.2.483-488.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  13. Merril C. R., Goldman D., Sedman S. A., Ebert M. H. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science. 1981 Mar 27;211(4489):1437–1438. doi: 10.1126/science.6162199. [DOI] [PubMed] [Google Scholar]
  14. Orlean P., Ammer H., Watzele M., Tanner W. Synthesis of an O-glycosylated cell surface protein induced in yeast by alpha factor. Proc Natl Acad Sci U S A. 1986 Sep;83(17):6263–6266. doi: 10.1073/pnas.83.17.6263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pierce M., Ballou C. E. Cell-cell recognition in yeast. Characterization of the sexual agglutination factors from Saccharomyces kluyveri. J Biol Chem. 1983 Mar 25;258(6):3576–3582. [PubMed] [Google Scholar]
  16. Sakai K., Yanagishima N. Mating reaction in Saccharomyces cerevisiae. II. Hormonal regulation of agglutinability of a type cells. Arch Mikrobiol. 1972;84(3):191–198. doi: 10.1007/BF00425197. [DOI] [PubMed] [Google Scholar]
  17. Shenbagamurthi P., Baffi R., Khan S. A., Lipke P., Pousman C., Becker J. M., Naider F. Structure-activity relationships in the dodecapeptide alpha factor of Saccharomyces cerevisiae. Biochemistry. 1983 Mar 1;22(5):1298–1304. doi: 10.1021/bi00274a047. [DOI] [PubMed] [Google Scholar]
  18. Sprague G. F., Jr, Blair L. C., Thorner J. Cell interactions and regulation of cell type in the yeast Saccharomyces cerevisiae. Annu Rev Microbiol. 1983;37:623–660. doi: 10.1146/annurev.mi.37.100183.003203. [DOI] [PubMed] [Google Scholar]
  19. Strazdis J. R., MacKay V. L. Reproducible and rapid methods for the isolation and assay of a-factor, a yeast mating hormone. J Bacteriol. 1982 Sep;151(3):1153–1161. doi: 10.1128/jb.151.3.1153-1161.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Taylor N. W., Orton W. L. Association constant of the sex-specific agglutinin in the yeast, Hansenula wingei. Biochemistry. 1970 Jul 7;9(14):2931–2934. doi: 10.1021/bi00816a027. [DOI] [PubMed] [Google Scholar]
  21. Taylor N. W., Orton W. L. Sexual agglutination in yeast. VII. Significance of the 1.7S component from reduced 5-agglutinin. Arch Biochem Biophys. 1968 Sep 10;126(3):912–921. doi: 10.1016/0003-9861(68)90485-2. [DOI] [PubMed] [Google Scholar]
  22. Terrance K., Lipke P. N. Sexual agglutination in Saccharomyces cerevisiae. J Bacteriol. 1981 Dec;148(3):889–896. doi: 10.1128/jb.148.3.889-896.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Wilkinson L. E., Pringle J. R. Transient G1 arrest of S. cerevisiae cells of mating type alpha by a factor produced by cells of mating type a. Exp Cell Res. 1974 Nov;89(1):175–187. doi: 10.1016/0014-4827(74)90200-6. [DOI] [PubMed] [Google Scholar]
  24. Yamaguchi M., Yoshida K., Yanagishima N. Isolation and partial characterization of cytoplasmic alpha agglutination substance in the yeast Saccharomyces cerevisiae. FEBS Lett. 1982 Mar 8;139(1):125–129. doi: 10.1016/0014-5793(82)80502-4. [DOI] [PubMed] [Google Scholar]
  25. Yoshida K., HAGIYA M., Yanagishima N. Isolation and purification of the sexual agglutination substance of mating type a cells in Saccharomyces cerevisiae. Biochem Biophys Res Commun. 1976 Aug 23;71(4):1085–1094. doi: 10.1016/0006-291x(76)90765-8. [DOI] [PubMed] [Google Scholar]

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