Two-dimensional gel analysis of yeast proteins: application to the study of changes in the levels of major polypeptides of Saccharomyces cerevisiae depending on the fermentable or nonfermentable nature of the carbon source.
Taking advantage of the recent identification of polypeptides of the carbon metabolism machinery on the yeast protein map [1], we applied two-dimensional gel electrophoresis to a study of changes in protein composition of Saccharomyces cerevisiae depending on the fermentable or nonfermentable nature of the carbon source. The levels of the 250 most abundant polypeptides were compared. Thirty-three were found to display markedly increased levels during growth on nonfermentable carbon sources. These 33 polypeptides include 11 mitochondrial polypeptides and polypeptides corresponding to alcohol dehydrogenase II, acetyl-CoA synthetase, phosphoenol pyruvate kinase and hexokinase PI. Sixteen other polypeptides, in contrast, reached their higher levels during growth on fermentable carbon sources. Among these were identified the monomeric subunits of 6 glycolytic enzymes. Collectively the 33 polypeptides of the first class comprised over 30% of the total soluble proteins of cells grown on nonfermentable carbon source and 3% during growth on fermentable carbon source. The protein fraction of the 16 polypeptides of the second class corresponded to 10% and 38%, respectively. Together these results show that two-dimensional gel electrophoresis, when coupled with the identification of polypeptides of the carbon metabolism apparatus, provides a valuable tool for approaching questions concerning carbon metabolism in S. cerevisiae.
Bataillé
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Laboratoire de Génetique, Faculté des Sciences, Talence, France.
Thoraval
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D
Boucherie
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H
D000085:Acetates / Q000378:metabolism; D002244:Carbon / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005285:Fermentation; D005656:Fungal Proteins / Q000032:analysis* / Q000096:biosynthesis; D005947:Glucose / Q000378:metabolism; D010449:Peptide Mapping; D012441:Saccharomyces cerevisiae / Q000032:analysis* / Q000378:metabolism
D000085:Acetates; D005656:Fungal Proteins; D002244:Carbon; D005947:Glucose