QuantiSpec — Quantitative mass spectrometry data analysis of N-metabolically labeled proteins |
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Authors: | Katrin Haegler Nikola S Mueller Giuseppina Maccarrone Eva Hunyadi-Gulyas Christian Webhofer Michaela D Filiou Yaoyang Zhang Christoph W Turck |
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Institution: | aMax Planck Institute of Psychiatry, Proteomics and Biomarkers, Munich, Germany |
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Abstract: | For relative protein quantitation by mass spectrometry we metabolically labeled E. coli bacteria with 15N-enriched diets. Proteins extracted from 15N-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 15N 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 14N/15N peptide pair. |
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Keywords: | Quantitative mass spectrometry Fast Fourier Transform Partial metabolic labeling Stable isotopes Bioinformatics Computer software |
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