Dataset features


Dataset type: Other
Number of samples: 1
Release date: Sep 26 2007
Access: Public
Chemicals: Amino Acids, Ammonium Sulfate, Carbon, Glucose, Nitrogen, Oxygen
Taxon: Saccharomyces cerevisiae
Dataset link Quantitative proteomics of anaerobic and aerobic yeast cultures

Experimental Protocol

The relative protein levels of anaerobically versus aerobically grown S. cerevisiae were investigated using metabolic stable isotope labeling. The biomass was produced using carbon limited chemostat cultures. Each biomass was produced twice, once using 14N labeled nitrogen source (ammonium sulfate) and once using 98% 15N nitrogen source. Subsequently, biomass was harvested, and protein extracts were prepared. Two mixed samples were prepared from these four protein samples, containing equal protein content of 14N aerobic and 15N anaerobic cultures for the first sample and equal protein content of 15N aerobic and 14N anaerobic cultures for the second sample. The two resulting samples were separated on an SDS-PAGE gel, and each lane was equally fragmented into 40 slices resulting in sub-samples based on protein size. This was followed by an in-gel protein digestion using trypsin, and analysis in triplicate by nanoflow-LC-MS/MS. Proteins were identified using MS/MS, and the relative protein levels were obtained by comparing the extracted ion chromatograms of the co-eluting 14N and 15N labeled peptide pairs