title: Lateral diffusion contributes to FRET from lanthanide-tagged membrane proteins.
abstract: Diffusion can enhance Förster resonance energy transfer (FRET) when donors or acceptors diffuse distances that are similar to the distances separating them during the donor's excited state lifetime. Lanthanide donors remain in the excited state for milliseconds, which makes them useful for time-resolved FRET applications but also allows time for diffusion to enhance energy transfer. Here we show that diffusion dramatically enhances FRET between membrane proteins labeled with lanthanide donors. This phenomenon complicates interpretation of experiments that use long-lived donors to infer association or proximity of mobile membrane proteins, but also offers a method of monitoring diffusion in membrane domains in real time in living cells.
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA.
authors.forename: Tien-Hung
authors.initials: TH
authors.lastname: Lan
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA.
authors.forename: Guangyu
authors.initials: G
authors.lastname: Wu
authors.affiliation: Department of Pharmacology and Toxicology, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA. Electronic address: nelambert@gru.edu.
authors.forename: Nevin A
authors.initials: NA
authors.lastname: Lambert
mesh_terms: D004058:Diffusion; D031541:Fluorescence Resonance Energy Transfer / Q000592:standards*; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D028581:Lanthanoid Series Elements / Q000737:chemistry* / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D050600:SNARE Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013194:Staining and Labeling
chemical_list: D028581:Lanthanoid Series Elements; D018343:Receptors, Adrenergic, beta-2; D011993:Recombinant Fusion Proteins; D050600:SNARE Proteins
keywords: Diffusion; FRET; GPCR; GPI-anchored proteins; HTRF; TR-FRET