Yeast Protein Complexes

Yeast Protein Complexes


Collaboration with the University of New South Wales Increasingly, proteomic projects are focusing on protein-protein interactions as a way of understanding how changes in a sample’s proteome affect the sample’s phenotype. Traditionally, two methods have been used to study protein-protein interactions, is the yeast two-hybrid system and the TAP-tag method. Both approaches have limitations and used in terms of a global approach, can be expensive and time consuming. PTCE staff in collaboration with Professor Mark Wilkins group at the University of New South Wales have been investigating the use of Blue Native PAGE in the Bio-Rad PrepCell (Blue Native Preparative Electrophoresis) to separate and purify protein complexes from crude cell extracts and then directly analyze the fractionated proteins with mass spectrometry to get a global impression of the protein-protein interaction network in yeast and determine the differences in protein complexes between phenotypically different yeast strains. We have been able to successfully separate, isolate and analyse protein complexes of 1000 kDa in size using Blue Native Preparative Electrophoresis and are now investigating the use of DIGE for the labeling of different yeast sample prior to mixing the samples and performing Blue Native Preparative Electrophoresis.