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Chromatographic isolation of methionine-containing peptides for gel-free proteome analysis: identification of more than 800 Escherichia coli proteins

Kris Gevaert UGent, Jozef Van Damme UGent, Marc Goethals UGent, Grégoire Thomas, Bart Hoorelbeke, Hans Demol UGent, Lennart Martens UGent, Magda Puype UGent, An Staes UGent and Joël Vandekerckhove UGent (2002) MOLECULAR & CELLULAR PROTEOMICS. 1(11). p.896-903
abstract
A novel gel-free proteomic technology was used to identify more than 800 proteins from 50 million Escherichia coli K12 cells in a single analysis. A peptide mixture is first obtained from a total unfractionated cell lysate, and only the methionine-containing peptides are isolated and identified by mass spectrometry and database searching. The sorting procedure is based on the concept of diagonal chromatography but adapted for highly complex mixtures. Statistical analysis predicts that we have identified more than 40% of the expressed proteome, including soluble and membrane-bound proteins. Next to highly abundant proteins, we also detected low copy number components such as the E. coli lactose operon repressor, illustrating the high dynamic range. The method is about 100 times more sensitive than two-dimensional gel-based methods and is fully automated. The strongest point, however, is the flexibility in the peptide sorting chemistry, which may target the technique toward quantitative proteomics of virtually every class of peptides containing modifiable amino acids, such as phosphopeptides, amino-terminal peptides, etc., adding a new dimension to future proteome research.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
SEQUENCE, DESORPTION/IONIZATION MASS-SPECTROMETRY, CODED AFFINITY TAGS, MIXTURES, PHOSPHOPROTEOME, ELECTROSPRAY
journal title
MOLECULAR & CELLULAR PROTEOMICS
Mol. Cell. Proteomics
volume
1
issue
11
pages
896 - 903
Web of Science type
Article
Web of Science id
000181516000005
ISSN
1535-9476
DOI
10.1074/mcp.M200061-MCP200
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
208762
handle
http://hdl.handle.net/1854/LU-208762
date created
2008-01-11 10:41:00
date last changed
2013-01-30 09:42:52
@article{208762,
  abstract     = {A novel gel-free proteomic technology was used to identify more than 800 proteins from 50 million Escherichia coli K12 cells in a single analysis. A peptide mixture is first obtained from a total unfractionated cell lysate, and only the methionine-containing peptides are isolated and identified by mass spectrometry and database searching. The sorting procedure is based on the concept of diagonal chromatography but adapted for highly complex mixtures. Statistical analysis predicts that we have identified more than 40\% of the expressed proteome, including soluble and membrane-bound proteins. Next to highly abundant proteins, we also detected low copy number components such as the E. coli lactose operon repressor, illustrating the high dynamic range. The method is about 100 times more sensitive than two-dimensional gel-based methods and is fully automated. The strongest point, however, is the flexibility in the peptide sorting chemistry, which may target the technique toward quantitative proteomics of virtually every class of peptides containing modifiable amino acids, such as phosphopeptides, amino-terminal peptides, etc., adding a new dimension to future proteome research.},
  author       = {Gevaert, Kris and Van Damme, Jozef and Goethals, Marc and Thomas, Gr{\'e}goire and Hoorelbeke, Bart and Demol, Hans and Martens, Lennart and Puype, Magda and Staes, An and Vandekerckhove, Jo{\"e}l},
  issn         = {1535-9476},
  journal      = {MOLECULAR \& CELLULAR PROTEOMICS},
  keyword      = {SEQUENCE,DESORPTION/IONIZATION MASS-SPECTROMETRY,CODED AFFINITY TAGS,MIXTURES,PHOSPHOPROTEOME,ELECTROSPRAY},
  language     = {eng},
  number       = {11},
  pages        = {896--903},
  title        = {Chromatographic isolation of methionine-containing peptides for gel-free proteome analysis: identification of more than 800 Escherichia coli proteins},
  url          = {http://dx.doi.org/10.1074/mcp.M200061-MCP200},
  volume       = {1},
  year         = {2002},
}

Chicago
Gevaert, Kris, Jozef Van Damme, Marc Goethals, Grégoire Thomas, Bart Hoorelbeke, Hans Demol, Lennart Martens, Magda Puype, An Staes, and Joël Vandekerckhove. 2002. “Chromatographic Isolation of Methionine-containing Peptides for Gel-free Proteome Analysis: Identification of More Than 800 Escherichia Coli Proteins.” Molecular & Cellular Proteomics 1 (11): 896–903.
APA
Gevaert, K., Van Damme, J., Goethals, M., Thomas, G., Hoorelbeke, B., Demol, H., Martens, L., et al. (2002). Chromatographic isolation of methionine-containing peptides for gel-free proteome analysis: identification of more than 800 Escherichia coli proteins. MOLECULAR & CELLULAR PROTEOMICS, 1(11), 896–903.
Vancouver
1.
Gevaert K, Van Damme J, Goethals M, Thomas G, Hoorelbeke B, Demol H, et al. Chromatographic isolation of methionine-containing peptides for gel-free proteome analysis: identification of more than 800 Escherichia coli proteins. MOLECULAR & CELLULAR PROTEOMICS. 2002;1(11):896–903.
MLA
Gevaert, Kris, Jozef Van Damme, Marc Goethals, et al. “Chromatographic Isolation of Methionine-containing Peptides for Gel-free Proteome Analysis: Identification of More Than 800 Escherichia Coli Proteins.” MOLECULAR & CELLULAR PROTEOMICS 1.11 (2002): 896–903. Print.