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  A proteomic method for the analysis of changes in protein concentrations in response to systemic perturbations using metabolic incorporation of stable isotopes and mass spectrometry

Gustavsson, N., Greber, B., Kreitler, T., Himmelbauer, H., Lehrach, H., & Gobom, J. (2005). A proteomic method for the analysis of changes in protein concentrations in response to systemic perturbations using metabolic incorporation of stable isotopes and mass spectrometry. PROTEOMICS, 5(14), 3563-3570. doi:10.1002/pmic.200401193.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-85D5-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-85D6-7
Genre: Journal Article

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Gustavsson et al. - Proteomics.pdf (Any fulltext), 552KB
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Gustavsson et al. - Proteomics.pdf
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 Creators:
Gustavsson, Niklas1, Author
Greber, Boris1, Author
Kreitler, Thomas1, Author
Himmelbauer, Heinz2, Author              
Lehrach, Hans2, Author              
Gobom, Johan2, Author              
Affiliations:
1Max Planck Society, ou_persistent13              
2Dept. of Vertebrate Genomics (Head: Hans Lehrach), Max Planck Institute for Molecular Genetics, Max Planck Society, ou_1433550              

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Free keywords: MS; PTM; Protein turnover; Quantitative proteomics; Stable isotope labeling
 Abstract: While several techniques exist for assessing quantitative differences among proteomes representing different cell states, methods for assessing how these differences are mediated are largely missing. We present a method that allows one to differentiate between cellular processes, such as protein synthesis, degradation and PTMs which affect protein concentrations. An induced systemic perturbation of a cell culture was coupled to a replacement of the growth medium to one highly enriched in the stable isotope 15N. The relative abundance of the 15N- and 14N-enriched forms of proteins, isolated from cell cultures harvested at time points following the onset of the perturbation, were determined by MS. Alterations in protein synthesis and degradation were quantified by comparing proteins isolated from perturbed and unperturbed cultures, respectively. The method was evaluated by subjecting HeLa cells to heat stress. As expected, a number of known heat shock proteins (Hsp) increased in concentration during heat stress. For Hsp27, increased de novo synthesis accounted for the concentration increase, while for Hsp70, decreased degradation accounted for the increase. A protein that was detected only after prolonged heat stress, vimentin, was not primarily synthesized de novo, but appeared rather as a result of PTM.

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Language(s): eng - English
 Dates: 2005-08-01
 Publication Status: Published in print
 Pages: -
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 Table of Contents: -
 Rev. Method: -
 Identifiers: eDoc: 271542
DOI: 10.1002/pmic.200401193
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Title: PROTEOMICS
Source Genre: Journal
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Pages: - Volume / Issue: 5 (14) Sequence Number: - Start / End Page: 3563 - 3570 Identifier: ISSN: 1615-9853