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Title: Advanced Mass Spectrometric Methods for the Rapid and Quantitative Characterization of Proteomes

Progress is reviewed towards the development of a global strategy that aims to extend the sensitivity, dynamic range, comprehensiveness and throughput of proteomic measurements based upon the use of high performance separations and mass spectrometry. The approach uses high accuracy mass measurements from Fourier transform ion cyclotron resonance mass spectrometry (FTICR) to validate peptide ‘accurate mass tags’ (AMTs) produced by global protein enzymatic digestions for a specific organism, tissue or cell type from ‘potential mass tags’ tentatively identified using conventional tandem mass spectrometry (MS/MS). This provides the basis for subsequent measurements without the need for MS/ MS. High resolution capillary liquid chromatography separations combined with high sensitivity, and high resolution accurate FTICR measurements are shown to be capable of characterizing peptide mixtures of more than 10 5 components. The strategy has been initially demonstrated using the microorganisms Saccharomyces cerevisiae and Deinococcus radiodurans. Advantages of the approach include the high confidence of protein identification, its broad proteome coverage, high sensitivity, and the capability for stableisotope labeling methods for precise relative protein abundance measurements. Abbreviations : LC, liquid chromatography; FTICR, Fourier transform ion cyclotron resonance; AMT, accurate mass tag; PMT, potential mass tag; MMA, mass measurement accuracy; MS, mass spectrometry; MS/MS, tandem mass spectrometry; ppm, parts per million.
Authors:
 [1]
  1. Environmental and Molecular Sciences Laboratory, MSIN: K8-98, Pacific Northwest National Laboratory, P.O. Box 999, Richland, WA 99352, USA
Publication Date:
OSTI Identifier:
1197925
Grant/Contract Number:
AC06-76RLO1830
Type:
Published Article
Journal Name:
Comparative and Functional Genomics
Additional Journal Information:
Journal Volume: 3; Journal Issue: 2; Related Information: CHORUS Timestamp: 2016-08-23 04:56:00; Journal ID: ISSN 1531-6912
Publisher:
Hindawi Publishing Corporation
Sponsoring Org:
USDOE
Country of Publication:
Country unknown/Code not available
Language:
English