| EJMS Protocol: An improved calibration method for the matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resononance analysis of (15)N-metabolically- labeled proteome digests using a mass difference approach. | |
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MedLine Citation:
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PMID: 22837438 Owner: NLM Status: In-Data-Review |
Abstract/OtherAbstract:
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High mass measurement accuracy of peptides in enzymatic digests is critical for confident protein identification and characterization in proteomics research. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) can provide low or sub-ppm mass accuracy and ultrahigh resolving power. While for ESI-FT-ICR-MS, the mass accuracy is generally 1 ppm or better, with matrix-assisted laser desorption/ionization (MALDI)-FT-ICR-MS, the mass errors can vary from sub-ppm with internal calibration to over 100 ppm with conventional external calibration. A novel calibration method for (15)N-metabolically labeled peptides from a batch digest of a proteome is described which corrects for space charge induced frequency shifts in FT-ICR spectra without using an internal calibrant. This strategy utilizes the information from the mass difference between the (14)N/(15)N peptide peak pairs to correct for space charge induced mass shifts after data collection. A procedure for performing the mass correction has been written into a computer program and has been successfully applied to high-performance liquid chromatography-MALDI-FT- ICR-MS measurement of (15)N-metabolic labeled proteomes. We have achieved an average measured mass error of 1.0 ppm and a standard deviation of 3.5 ppm for 900 peptides from 68 MALDI-FT-ICR mass spectra of the proteolytic digest of a proteome from Methanococcus maripaludis. |
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Authors:
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Li Jing; I Jonathan Amster |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: European journal of mass spectrometry (Chichester, England) Volume: 18 ISSN: 1469-0667 ISO Abbreviation: Eur J Mass Spectrom (Chichester, Eng) Publication Date: 2012 |
Date Detail:
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Created Date: 2012-07-27 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101124748 Medline TA: Eur J Mass Spectrom (Chichester, Eng) Country: England |
Other Details:
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Languages: eng Pagination: 269-77 Citation Subset: IM |
Affiliation:
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Protea BioSciences, Inc., Morgantown, WV 26507, USA. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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