title: Analyzing transmembrane protein and hydrophobic helix topography by dual fluorescence quenching.
abstract: The location of fluorescent groups relative to the lipid bilayer can be evaluated using fluorescence quenchers embedded in the membrane and/or dissolved in aqueous solution. Quenching can be used to define the membrane topography of membrane proteins and individual membrane-embedded hydrophobic helices by combining it with the placement of fluorescent groups, including Trp, at defined sequence positions. This chapter briefly discusses various quenching methods for studies of membrane protein topography and provides detailed protocols for dual quencher analysis, a rapid, highly sensitive, and experimentally flexible approach in which the information gained from both a membrane-embedded and aqueous quencher is combined.
authors: Department of Chemistry and Biochemistry, Rowan University, Glassboro, NJ, USA.
authors: Gregory A
authors: GA
authors: Caputo
authors: Erwin
authors: E
authors: London
mesh_terms: D057927:Hydrophobic and Hydrophilic Interactions; D008051:Lipid Bilayers / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000378:metabolism*; D017433:Protein Structure, Secondary; D012996:Solutions; D013050:Spectrometry, Fluorescence
chemical_list: D008051:Lipid Bilayers; D008565:Membrane Proteins; D012996:Solutions