title: Green fluorescent protein expression triggers proteome changes in breast cancer cells.
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.
authors.affiliation: School of Science and Technology, University of New England, Armidale, NSW, Australia; School of Rural Medicine, University of New England, Armidale, NSW, Australia. Electronic address: jmoensco@une.edu.au.
authors.forename: J V F
authors.initials: JV
authors.lastname: Coumans
authors.forename: D
authors.initials: D
authors.lastname: Gau
authors.forename: A
authors.initials: A
authors.lastname: Poljak
authors.forename: V
authors.initials: V
authors.lastname: Wasinger
authors.forename: P
authors.initials: P
authors.lastname: Roy
authors.forename: P
authors.initials: P
authors.lastname: Moens
mesh_terms: D016477:Artifacts; D001943:Breast Neoplasms / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020543:Proteome / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D020543:Proteome; D049452:Green Fluorescent Proteins
keywords: 2-DE; Cellular immune response; Cytoskeleton modifications; GFP; GFP expression; Proteomic change; Unfolded protein response; green fluorescence protein; iTRAQ; isobaric tags for relative and absolute quantitation; two-dimensional electrophoresis
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