title: Quantitative measurement of intracellular protein dynamics using photobleaching or photoactivation of fluorescent proteins.
abstract: Unlike in vitro protein dynamics, intracellular protein dynamics are intricately regulated by protein-protein interactions or interactions between proteins and other cellular components, including nucleic acids, the plasma membrane and the cytoskeleton. Alteration of these dynamics plays a crucial role in physiological phenomena such as gene expression and cell division. Live-cell imaging via microscopy with the inherent properties of fluorescent proteins, i.e. photobleaching and photoconversion, or fluorescence correlation spectroscopy, provides insight into the movement of proteins and their interactions with cellular components. This article reviews techniques based on photo-induced changes in the physicochemical properties of fluorescent proteins to measure protein dynamics inside living cells, and it also discusses the strengths and weaknesses of these techniques.
authors.affiliation: Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan.
authors.forename: Tomoki
authors.initials: T
authors.lastname: Matsuda
authors.affiliation: Institute of Scientific and Industrial Research, Osaka University, 8-1 Mihogaoka, Ibaraki, Osaka 567-0047, Japan ng1@sanken.osaka-u.ac.jp.
authors.forename: Takeharu
authors.initials: T
authors.lastname: Nagai
mesh_terms: D002477:Cells / Q000737:chemistry* / Q000378:metabolism; D036681:Fluorescence Recovery After Photobleaching / Q000379:methods; D049452:Green Fluorescent Proteins; D008856:Microscopy, Fluorescence / Q000379:methods; D038761:Photobleaching; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D011506:Proteins; D049452:Green Fluorescent Proteins
keywords: FDAP; FRAP; diffusion coefficient; fluorescent protein; photoactivation; photobleaching