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Title: High throughput proteome-wide precision measurements of protein expression using mass spectrometry

Abstract

In contrast to a cell's virtually static genome, the proteome, the protein complement expressed by an organism, continually changes in response to external stimuli and internal processes. Global gene expression analysis at the mRNA level (i.e., transcriptome) has recently become feasible based on the serial analysis of gene expression and oligonucleotide micro-array assays. These techniques allow the activation states of thousands of genes to be polled simultaneously for a tissue or cell population. However, assays that measure mRNA abundances rather than the functional gene products (i.e., proteins) are uninformative with regard to protein modifications, and can poorly reflect protein abundances due to differences in stabilities, expression rates, etc., for both the mRNAs and proteins. The authors have developed an approach utilizing organisms cultured in stable-isotope labeled media (e.g., rare-isotope depleted and normal) to provide effective internal calibrants for all detected proteins, thus enabling precise proteome-wide measurement of changes in protein abundances resulting from cellular perturbations. The two (or more) isotopically distinctive cell populations are mixed prior to sample processing steps, eliminating all experimental variables associated with cell lysis, separation, and mass spectrometric analysis. Changes in relative protein abundances are thus precisely reflected by the ratio of two isotopically different andmore » resolvable versions of each protein.« less

Authors:
; ; ; ; ; ; ; ;
Publication Date:
Research Org.:
Environmental Molecular Sciences Lab., Richland, WA (US)
Sponsoring Org.:
USDOE
OSTI Identifier:
20000033
DOE Contract Number:  
AC06-76RL01830
Resource Type:
Journal Article
Journal Name:
Journal of the American Chemical Society
Additional Journal Information:
Journal Volume: 121; Journal Issue: 34; Other Information: PBD: 1 Sep 1999; Journal ID: ISSN 0002-7863
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; PROTEINS; STABLE ISOTOPES; LABELLED COMPOUNDS; ABUNDANCE; ISOTOPE DILUTION; ION CYCLOTRON RESONANCE SPECTROSCOPY; FOURIER TRANSFORM SPECTROMETERS

Citation Formats

Pasa-Tolic, L., Jensen, P.K., Anderson, G.A., Lipton, M.S., Peden, K.K., Martinovic, S., Tolic, N., Bruce, J.E., and Smith, R.D. High throughput proteome-wide precision measurements of protein expression using mass spectrometry. United States: N. p., 1999. Web. doi:10.1021/ja991063o.
Pasa-Tolic, L., Jensen, P.K., Anderson, G.A., Lipton, M.S., Peden, K.K., Martinovic, S., Tolic, N., Bruce, J.E., & Smith, R.D. High throughput proteome-wide precision measurements of protein expression using mass spectrometry. United States. doi:10.1021/ja991063o.
Pasa-Tolic, L., Jensen, P.K., Anderson, G.A., Lipton, M.S., Peden, K.K., Martinovic, S., Tolic, N., Bruce, J.E., and Smith, R.D. Wed . "High throughput proteome-wide precision measurements of protein expression using mass spectrometry". United States. doi:10.1021/ja991063o.
@article{osti_20000033,
title = {High throughput proteome-wide precision measurements of protein expression using mass spectrometry},
author = {Pasa-Tolic, L. and Jensen, P.K. and Anderson, G.A. and Lipton, M.S. and Peden, K.K. and Martinovic, S. and Tolic, N. and Bruce, J.E. and Smith, R.D.},
abstractNote = {In contrast to a cell's virtually static genome, the proteome, the protein complement expressed by an organism, continually changes in response to external stimuli and internal processes. Global gene expression analysis at the mRNA level (i.e., transcriptome) has recently become feasible based on the serial analysis of gene expression and oligonucleotide micro-array assays. These techniques allow the activation states of thousands of genes to be polled simultaneously for a tissue or cell population. However, assays that measure mRNA abundances rather than the functional gene products (i.e., proteins) are uninformative with regard to protein modifications, and can poorly reflect protein abundances due to differences in stabilities, expression rates, etc., for both the mRNAs and proteins. The authors have developed an approach utilizing organisms cultured in stable-isotope labeled media (e.g., rare-isotope depleted and normal) to provide effective internal calibrants for all detected proteins, thus enabling precise proteome-wide measurement of changes in protein abundances resulting from cellular perturbations. The two (or more) isotopically distinctive cell populations are mixed prior to sample processing steps, eliminating all experimental variables associated with cell lysis, separation, and mass spectrometric analysis. Changes in relative protein abundances are thus precisely reflected by the ratio of two isotopically different and resolvable versions of each protein.},
doi = {10.1021/ja991063o},
journal = {Journal of the American Chemical Society},
issn = {0002-7863},
number = 34,
volume = 121,
place = {United States},
year = {1999},
month = {9}
}