title: The substituted-cysteine accessibility method (SCAM) to elucidate membrane protein structure.
abstract: The substituted-cysteine accessibility method (SCAM) provides an approach to identifying the residues in the membrane-spanning segments that line a channel, transporter, or binding-site crevice. SCAM can also be used to determine differences in the structures of the membrane-spanning segments in different functional states of the proteins, to map electrostatic potential in the membrane-spanning domains, and to size a channel or binding-site crevice. The protocol in this unit describes the use of SCAM to map the binding-site crevice of a G-protein coupled receptor (GPCR) which binds ligand within the transmembrane portion of the receptor.
authors.affiliation: Columbia University, New York, New York, USA.
authors.forename: G
authors.initials: G
authors.lastname: Liapakis
authors.forename: M M
authors.initials: MM
authors.lastname: Simpson
authors.forename: J A
authors.initials: JA
authors.lastname: Javitch
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics*; D001665:Binding Sites / Q000235:genetics; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry / Q000235:genetics; D002874:Chromosome Mapping / Q000379:methods*; D003545:Cysteine / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics*; D009154:Mutation; D013329:Structure-Activity Relationship; D013439:Sulfhydryl Reagents / Q000737:chemistry
chemical_list: D008565:Membrane Proteins; D013439:Sulfhydryl Reagents; D003545:Cysteine