Reagents for investigating MAPK signalling in model yeast species

Yeast. 2010 Jul;27(7):423-30. doi: 10.1002/yea.1758.


Here we present a set of resources (bacterial expression plasmids and antibodies) for the interrogation of proteins involved in yeast MAPK signalling. We constructed bacterial protein expression plasmids for 25 proteins involved in MAPK signalling in budding yeast. From these constructs we expressed and purified proteins and generated rabbit polyclonal antibodies against 13 proteins in the pheromone MAPK pathway. We verified the specificity of the antibodies and employed them to follow pathway proteins in cells stimulated with pheromone. We show that these reagents can be used to detect pheromone-induced post-translational modifications and changes in the oligomeric state of pathway proteins. In addition to recognizing their target proteins in Saccharomyces cerevisiae, these antibodies allow the detection of predicted orthologues in the distant evolutionary relatives Kluyveromyces lactis and Schizosaccharomyces pombe. These antibodies are new tools for investigating MAPK signalling in model yeast species and may be useful for studying MAPK signalling in higher eukaryotes.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Antibodies, Fungal / immunology*
  • Antibodies, Fungal / isolation & purification
  • Fungal Proteins / genetics
  • Fungal Proteins / isolation & purification
  • Fungal Proteins / metabolism*
  • Gene Expression
  • Gene Expression Regulation, Fungal*
  • Genetic Vectors
  • Kluyveromyces / metabolism
  • Kluyveromyces / physiology
  • Mitogen-Activated Protein Kinases / immunology
  • Mitogen-Activated Protein Kinases / metabolism*
  • Pheromones*
  • Plasmids
  • Protein Processing, Post-Translational
  • Rabbits
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces / physiology
  • Signal Transduction*
  • Yeasts / metabolism
  • Yeasts / physiology*


  • Antibodies, Fungal
  • Fungal Proteins
  • Pheromones
  • Recombinant Proteins
  • Mitogen-Activated Protein Kinases