Green fluorescent protein expression triggers proteome changes in breast cancer cells |
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| Authors: | J.V.F. Coumans D. Gau A. Poljak V. Wasinger P. Roy P. Moens |
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| Affiliation: | 1. School of Science and Technology, University of New England, Armidale, NSW, Australia;2. School of Rural Medicine, University of New England, Armidale, NSW, Australia;3. Bioengineering and Pathology, University of Pittsburgh, Pittsburgh, PA, United States;4. Bioanalytical Mass Spectrometry Facility, The University of New South Wales, Sydney, NSW, Australia;5. School of Medical Sciences, The University of New South Wales, Sydney, NSW, Australia |
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| Abstract: | Green fluorescent protein (GFP) is the most commonly used reporter of expression in cell biology despite evidence that it affects the cell physiology. The molecular mechanism of GFP-associated modifications has been largely unexplored. In this paper we investigated the proteome modifications following stable expression of GFP in breast cancer cells (MDA-MB-231). A combination of three different proteome analysis methods (2-DE, iTRAQ, label-free) was used to maximise proteome coverage. We found that GFP expression induces changes in expression of proteins that are associated with protein folding, cytoskeletal organisation and cellular immune response. In view of these findings, the use of GFP as a cell reporter should be carefully monitored. |
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| Keywords: | GFP, green fluorescence protein 2-DE, two-dimensional electrophoresis iTRAQ, isobaric tags for relative and absolute quantitation |
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