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A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates

(2009) PROTEOMICS. 9(3). p.598-609
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Abstract
While protein interaction studies and protein network modeling come to the forefront, the isolation and identification of protein complexes in a cellular context remains a major challenge for plant science. To this end, a nondenaturing extraction procedure was optimized for plant whole cell matrices and the combined use of gel filtration and BN-PAGE for the separation of protein complexes was studied. Hyphenation to denaturing electrophoresis and mass spectrometric analysis allows for the simultaneous identification of multiple (previously unidentified) protein interactions in single samples. The reliability and efficacy of the technique was confirmed (i) by the identification of well-studied plant protein complexes, (ii) by the presence of nonplant interologs for several of the novel complexes (iii) by presenting physical evidence of previously hypothetical plant protein interactions and (iv) by the confirmation of found interactions using co-IP. Furthermore practical issues concerning the use of this 2-D BN/SDS-PAGE display method for the analysis of protein-protein interactions are discussed.
Keywords
ARABIDOPSIS-THALIANA, AFFINITY PURIFICATION, MASS, IDENTIFICATION, CHLOROPLASTS, ISOFORMS, DIPHOSPHATE KINASE, BLUE NATIVE ELECTROPHORESIS, CYTOSOLIC GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE, POLYACRYLAMIDE-GEL ELECTROPHORESIS, Protein complexes, Blue native gel electrophoresis, Nicotiana tabacum cv. bright yellow-2

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Citation

Please use this url to cite or link to this publication:

Chicago
Remmerie, Noor, Luc Roef, Eveline Van De Slijke, Jelle Van Leene, Geert Persiau, Dominique Eeckhout, Hilde Stals, et al. 2009. “A Bioanalytical Method for the Proteome Wide Display and Analysis of Protein Complexes from Whole Plant Cell Lysates.” Proteomics 9 (3): 598–609.
APA
Remmerie, N., Roef, L., Van De Slijke, E., Van Leene, J., Persiau, G., Eeckhout, D., Stals, H., et al. (2009). A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates. PROTEOMICS, 9(3), 598–609.
Vancouver
1.
Remmerie N, Roef L, Van De Slijke E, Van Leene J, Persiau G, Eeckhout D, et al. A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates. PROTEOMICS. 2009;9(3):598–609.
MLA
Remmerie, Noor, Luc Roef, Eveline Van De Slijke, et al. “A Bioanalytical Method for the Proteome Wide Display and Analysis of Protein Complexes from Whole Plant Cell Lysates.” PROTEOMICS 9.3 (2009): 598–609. Print.
@article{528497,
  abstract     = {While protein interaction studies and protein network modeling come to the forefront, the isolation and identification of protein complexes in a cellular context remains a major challenge for plant science. To this end, a nondenaturing extraction procedure was optimized for plant whole cell matrices and the combined use of gel filtration and BN-PAGE for the separation of protein complexes was studied. Hyphenation to denaturing electrophoresis and mass spectrometric analysis allows for the simultaneous identification of multiple (previously unidentified) protein interactions in single samples. The reliability and efficacy of the technique was confirmed (i) by the identification of well-studied plant protein complexes, (ii) by the presence of nonplant interologs for several of the novel complexes (iii) by presenting physical evidence of previously hypothetical plant protein interactions and (iv) by the confirmation of found interactions using co-IP. Furthermore practical issues concerning the use of this 2-D BN/SDS-PAGE display method for the analysis of protein-protein interactions are discussed.},
  author       = {Remmerie, Noor and Roef, Luc and Van De Slijke, Eveline and Van Leene, Jelle and Persiau, Geert and Eeckhout, Dominique and Stals, Hilde and Laukens, Kris and Lemi{\`e}re, Filip and Esmans, Eddy and Van Onckelen, Harry and Inz{\'e}, Dirk and De Jaeger, Geert and Witters, Erwin},
  issn         = {1615-9853},
  journal      = {PROTEOMICS},
  keyword      = {ARABIDOPSIS-THALIANA,AFFINITY PURIFICATION,MASS,IDENTIFICATION,CHLOROPLASTS,ISOFORMS,DIPHOSPHATE KINASE,BLUE NATIVE ELECTROPHORESIS,CYTOSOLIC GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE,POLYACRYLAMIDE-GEL ELECTROPHORESIS,Protein complexes,Blue native gel electrophoresis,Nicotiana tabacum cv. bright yellow-2},
  language     = {eng},
  number       = {3},
  pages        = {598--609},
  title        = {A bioanalytical method for the proteome wide display and analysis of protein complexes from whole plant cell lysates},
  url          = {http://dx.doi.org/10.1002/pmic.200800100},
  volume       = {9},
  year         = {2009},
}

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