title: Direct iterative protein profiling (DIPP) - an innovative method for large-scale protein detection applied to budding yeast mitosis.
abstract: The budding yeast Saccharomyces cerevisiae is a major model organism for important biological processes such as mitotic growth and meiotic development, it can be a human pathogen, and it is widely used in the food-, and biotechnology industries. Consequently, the genomes of numerous strains have been sequenced and a very large amount of RNA profiling data is available. Moreover, it has recently become possible to quantitatively analyze the entire yeast proteome; however, efficient and cost-effective high-throughput protein profiling remains a challenge. We report here a new approach to direct and label-free large-scale yeast protein identification using a tandem buffer system for protein extraction, two-step protein prefractionation and enzymatic digestion, and detection of peptides by iterative mass spectrometry. Our profiling study of diploid cells undergoing rapid mitotic growth identified 86% of the known proteins and its output was found to be widely concordant with genome-wide mRNA concentrations and DNA variations between yeast strains. This paves the way for comprehensive and straightforward yeast proteome profiling across a wide variety of experimental conditions.
authors: Inserm U1085, IRSET, Proteomics Core Facility Biogenouest, Université de Rennes 1, F-35042 Rennes, France.
authors: Régis
authors: R
authors: Lavigne
authors: Emmanuelle
authors: E
authors: Becker
authors: Yuchen
authors: Y
authors: Liu
authors: Bertrand
authors: B
authors: Evrard
authors: Aurélie
authors: A
authors: Lardenois
authors: Michael
authors: M
authors: Primig
authors: Charles
authors: C
authors: Pineau
mesh_terms: D002853:Chromatography, Liquid; D004271:DNA, Fungal / Q000032:analysis / Q000235:genetics; D020869:Gene Expression Profiling; D006801:Humans; D008938:Mitosis / Q000502:physiology*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D004271:DNA, Fungal; D020543:Proteome; D029701:Saccharomyces cerevisiae Proteins