title: Functional Site-Directed Fluorometry.
abstract: Initially developed in the mid-1990s to examine the conformational changes of the canonical Shaker voltage-gated potassium channel, functional site-directed fluorometry has since been expanded to numerous other voltage-gated and ligand-gated ion channels as well as transporters, pumps, and other integral membrane proteins. The power of functional site-directed fluorometry, also known as voltage-clamp fluorometry, lies in its ability to provide information on the conformational changes in a protein in response to changes in its environment with high temporal resolution while simultaneously monitoring the function of that protein. Over time, applications of site-directed fluorometry have expanded to examine the interactions of ion channels with modulators ranging from membrane potential to ligands to accessory protein subunits to lipids. In the future, the range of questions answerable by functional site-directed fluorometry and its interpretive power should continue to improve, making it an even more powerful technique for dissecting the conformational dynamics of ion channels and other membrane proteins.
authors.affiliation: Department of Biochemistry and Molecular Biology and Committee on Neurobiology, University of Chicago, Gordon Center for Integrative Science W229M, 929 East 57th Street, 60637, Chicago, IL, USA. priestm@uchicago.edu.
authors.forename: Michael
authors.initials: M
authors.lastname: Priest
authors.affiliation: Department of Biochemistry and Molecular Biology and Committee on Neurobiology, University of Chicago, Gordon Center for Integrative Science W229M, 929 East 57th Street, 60637, Chicago, IL, USA. fbezanilla@uchicago.edu.
authors.forename: Francisco
authors.initials: F
authors.lastname: Bezanilla
mesh_terms: D000818:Animals; D001665:Binding Sites; D005470:Fluorometry / Q000379:methods*; D006801:Humans; D015640:Ion Channel Gating; D007473:Ion Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008564:Membrane Potentials; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011485:Protein Binding; D011487:Protein Conformation; D013329:Structure-Activity Relationship
chemical_list: D007473:Ion Channels; D008024:Ligands
keywords: Conformational change; Conformational dynamics; Fluorescent dye; Ligand-gated ion channels; Site-directed fluorometry; Voltage-clamp fluorometry; Voltage-gated ion channels