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

Smith, Richard D.

doi:10.1002/cfg.159

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

Journal Article · Tue Jan 01 00:00:00 EST 2002 · Comparative and Functional Genomics

DOI:https://doi.org/10.1002/cfg.159· OSTI ID:1197925

Smith, Richard D. ^[1]

Environmental and Molecular Sciences Laboratory, MSIN: K8-98, Pacific Northwest National Laboratory, P.O. Box 999, Richland, WA 99352, USA

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.

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Research Organization:: Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER). Biological Systems Science Division; National Institutes of Health (NIH); Eunice Kennedy Shriver National Institute for Child Health & Human Development (NICHD) National Cancer Institute (NCI); National Institute of Neurological Disorders and Stroke (NINDS); National Center for Research Resources (NCRR)

Grant/Contract Number:: AC06-76RL01830; AC05-76RL01830

OSTI ID:: 1197925

Alternate ID(s):: OSTI ID: 1623502

Journal Information:: Comparative and Functional Genomics, Journal Name: Comparative and Functional Genomics Vol. 3 Journal Issue: 2; ISSN 1531-6912

Publisher:: Hindawi Publishing CorporationCopyright Statement

Country of Publication:: Country unknown/Code not available

Language:: English

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Cited by: 36 works

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References (14)

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59 BASIC BIOLOGICAL SCIENCES
biochemistry & molecular biology
biotechnology & applied microbiology
genetics & heredity
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proteome
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D. radiodurans

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