期刊
JOURNAL OF PROTEOMICS
卷 71, 期 6, 页码 601-608出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jprot.2008.10.004
关键词
Quantitative mass spectrometry; Fast Fourier Transform; Partial metabolic labeling; Stable isotopes; Bioinformatics; Computer software
资金
- BMBF QuantPro
- Max Planck Society
For relative protein quantitation by mass spectrometry we metabolically labeled E. coli bacteria with N-15-enriched diets. Proteins extracted from N-15-labeled and unlabeled E. coli bacteria were mixed, separated by two-dimensional gel electrophoresis and enzymatically digested. The resulting tryptic peptides were analyzed by MALDI mass spectrometry. For the relative protein quantitation we developed fully automated software, QuantiSpec (Quantitative Mass Spectrometry Analysis Software), which uses data from MALDI TOF mass spectrometry and the Mascot database search engine. QuantiSpec detects natural as well as partially or fully labeled peptide isotope distributions. For each identified peptide the N-15 incorporation rate is determined by comparing the experimental to a set of theoretical isotope patterns based on the peptide sequence. Relative quantitation is accomplished by calculating the signal intensity ratios for each N-14/N-15 peptide pair. (C) 2008 Elsevier B.V. All rights reserved.
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