title: Analysis of in vivo ROP GTPase activity at the subcellular level by fluorescence resonance energy transfer microscopy.
abstract: Proteins generally interact with some other proteins to achieve their cellular functions. Fluorescence resonance energy transfer (FRET) microscopy provides a powerful technique to elucidate such interactions in vivo. FRET occurs when two properly chosen fluorophores are sufficiently close (less than 10 nm). Aided by multiple colored fluorescent proteins (FPs), FRET microscopy has been widely used in live cells for detection of protein-protein interaction and in some cases protein activity in a real-time in vivo manner, which contributes to the understanding of the mechanisms for the regulation of many cellular activities, such as signal transduction pathways. Here, we describe a convenient and fast FRET imaging microscopy involving transiently expressed proteins fused with an FRET pair of fluorescent proteins (e.g., cyn fluorescent protein and yellow fluorescent protein). We describe an example of the FRET-based assay used to analyze ROP GTPase activity in live plant cells.
authors.affiliation: State Key Laboratory of Plant Physiology and Biochemistry, Department of Plant Sciences, College of Biological Sciences, China Agricultural University, Beijing, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhu
authors.forename: Ying
authors.initials: Y
authors.lastname: Fu
mesh_terms: D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D008164:Luminescent Proteins / Q000378:metabolism*; D010940:Plant Proteins / Q000378:metabolism*; D011485:Protein Binding
chemical_list: D008164:Luminescent Proteins; D010940:Plant Proteins; D019204:GTP-Binding Proteins