Two-dimensional analysis of proteins secreted by Saccharomyces cerevisiae regenerating protoplasts: a novel approach to study the cell wall

Yeast. 1999 Apr;15(6):459-72. doi: 10.1002/(SICI)1097-0061(199904)15:6<459::AID-YEA387>3.0.CO;2-L.

Abstract

Protoplasts of Saccharomyces cerevisiae incubated in regenerating conditions secrete cell wall components in order to allow the biosynthesis of this structure. During the first hours of incubation, many of these are not retained in the forming cell wall but remain in the medium. We have developed a method for collecting the secreted proteins and have analysed these by two-dimensional electrophoresis to obtain a reference map of putative cell wall proteins. Several proteins were identified by microsequencing or immunoblotting; namely, cell wall hydrolytic enzymes, heat shock proteins, glycolytic enzymes and others. Some beta-1,3- and beta-1, 6-glucosylation was detected in the proteins secreted by regenerating protoplasts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Blotting, Western
  • Cell Wall / chemistry*
  • Cell Wall / enzymology
  • Cell Wall / metabolism
  • Cell Wall / physiology
  • Culture Media, Conditioned
  • Electrophoresis, Gel, Two-Dimensional
  • Fungal Proteins / analysis*
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism
  • Genes, Fungal
  • Genetic Complementation Test
  • Glucuronidase / metabolism
  • Glycoside Hydrolases / genetics
  • Glycosylation
  • Heat-Shock Proteins / analysis
  • Heat-Shock Proteins / isolation & purification
  • Heat-Shock Proteins / metabolism
  • Molecular Weight
  • Multienzyme Complexes / metabolism
  • Mutation
  • Protoplasts / metabolism*
  • Protoplasts / physiology
  • Protoplasts / ultrastructure
  • Saccharomyces cerevisiae / cytology*
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism
  • Sequence Analysis
  • Sucrose / metabolism
  • Sulfatases / metabolism
  • beta-Fructofuranosidase

Substances

  • Culture Media, Conditioned
  • Fungal Proteins
  • Heat-Shock Proteins
  • Multienzyme Complexes
  • glusulase
  • Sucrose
  • Sulfatases
  • Glycoside Hydrolases
  • beta-Fructofuranosidase
  • Glucuronidase