title: Analysis of protein dynamics with tandem fluorescent protein timers.
abstract: Fluorescent timers (FTs) are fluorescent proteins that change color with time. FTs can be used as tags to follow protein dynamics in living cells. Recently we described a novel class of FTs called tandem fluorescent protein timers (tFTs). Each tFT is a tandem fusion of two different conventional fluorescent proteins having distinct kinetics of fluorophore maturation. tFTs suitable for studying protein dynamics on different scales can be generated from a broad range of commonly used fluorescent proteins. Here we describe how to establish new tFTs and consider potential pitfalls. We detail a protocol for quantitative fluorescence microscopy imaging and analysis of intracellular protein dynamics with tFTs in the budding yeast Saccharomyces cerevisiae.
authors.affiliation: Zentrum für Molekulare Biologie der Universität Heidelberg, DKFZ-ZMBH Alliance, Im Neuenheimer Feld 282, Heidelberg, 69120, Germany.
authors.forename: Anton
authors.initials: A
authors.lastname: Khmelinskii
authors.forename: Michael
authors.initials: M
authors.lastname: Knop
mesh_terms: D042541:Intracellular Space; D008164:Luminescent Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence / Q000379:methods; D021381:Protein Transport; D011506:Proteins / Q000378:metabolism*; D011993:Recombinant Fusion Proteins; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D011506:Proteins; D011993:Recombinant Fusion Proteins