Archives of Microbiology

, Volume 148, Issue 3, pp 208–212

Isolation and composition of the constitutive agglutinins from haploid Saccharomyces cerevisiae cells

  • P. C. Sijmons
  • A. J. A. Nederbragt
  • F. M. Klis
  • H. Van Den Ende
Original Papers

DOI: 10.1007/BF00414813

Cite this article as:
Sijmons, P.C., Nederbragt, A.J.A., Klis, F.M. et al. Arch. Microbiol. (1987) 148: 208. doi:10.1007/BF00414813

Abstract

Sex-specific agglutinins from the cell surface of haploid cells of Saccharomyces cerevisiae (X2180, mta and mtα) were purified and analysed. The constitutive agglutinin from mta cells was extractable with 3 mM dithiothreitol. It was shown to be a glycoprotein (3% mannose) with an apparent Mr of 43,000 based on gel filtration, but in SDS-PAGE it behaved as a much smaller molecule (Mr between 18,000 and 26,000). About one in three amino acids was a hydroxyamino acid. Its biological activity was resistant to boiling for 1 h, but sensitive to pronase. Intact mtα cells retained their agglutinability in the presence of dithiothreitol but limited trypsinizing released a biologically active agglutinin fragment. It had an apparent Mr of 320,000 (gel filtration). When analysed by SDS-PAGE, a single diffuse band with an apparent Mr of 225,000 was observed. The protein was 94% (w/w) mannose with a trace of N-acetyl glucosamine. Its biological activity was almost completely lost after boiling for 1 h. Both agglutinins behaved as monovalent molecules and specifically inhibited the biological activity of both noninduced and pheromone-induced cells. Pheromone treatment of mta cells resulted in an apparent 32-fold increase in agglutinin activity at the cell surface, whereas pheromone treatment of mtα cells only doubled the apparent agglutinin activity.

Key words

Saccharomyces cerevisiae Yeast mating Cell-cell recognition Sexual agglutination Agglutinins 

Abbreviations

mt

mating type

DTT

dithiothreitol

SDS-PAGE

sodium dodecyl sulphate polyacrylamide gel electrophoresis

YPG

yeast-peptone-glucose

PAS

periodic-acid-Schiff reagent

Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • P. C. Sijmons
    • 1
  • A. J. A. Nederbragt
    • 1
  • F. M. Klis
    • 1
  • H. Van Den Ende
    • 1
  1. 1.Department of Plant Physiology, Biotechnology CenterUniversity of AmsterdamAmsterdamThe Netherlands
  2. 2.MOGEN International BVLeidenThe Netherlands