title: Photoactivatable-GFP-α-tubulin as a tool to study microtubule plus-end turnover in living human cells.
abstract: The development of photactivatable (PA) variants of Green fluorescent protein (GFP) has allowed the dynamics of spatially restricted protein pools within living cells to be determined. Over the last 5 years, experiments utilizing PA-GFP fused to α-tubulin have provided important insights into the mechanisms that control microtubule dynamics in living cells. In this chapter, we describe the methodology required to generate stable cell lines expressing photoactivatable-GFP-α-tubulin and to derive quantitative measurements of tubulin turnover at microtubules plus-ends in living cells.
authors.affiliation: Centre for Mechanochemical Cell Biology, Warwick Medical School, University of Warwick, Coventry, UK. catarina@mechanochemistry.org
authors.forename: Catarina P
authors.initials: CP
authors.lastname: Samora
authors.forename: Andrew D
authors.initials: AD
authors.lastname: McAinsh
mesh_terms: D002460:Cell Line; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008870:Microtubules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014404:Tubulin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D011993:Recombinant Fusion Proteins; D014404:Tubulin; D049452:Green Fluorescent Proteins