Cysteine mutants as chemical sensors for ligand-receptor interactions.
The incorporation of cysteine residues into membrane receptors by mutagenesis has enabled the development of engineered proteins. Chemical modification of the mutant receptor using a wide range of biochemical and biophysical probes has facilitated functional studies of ligand-receptor interactions. In particular, the substituted-cysteine accessibility method (SCAM) represents a successful example of how to probe transmembrane receptor domains after chemical modification of the mutants with sulfydryl-reacting molecules. We propose an extension of this methodology using site-specific affinity probes that react with cysteine mutants to gain reliable structural information on the binding of a ligand in its receptor site.
Laboratoire de Chimie Bioorganique, UMR 7514 CNRS, Faculté de Pharmacie, Université Louis Pasteur Strasbourg, BP 24, 67401 ILLKIRCH, Cedex, France.
B
B
Foucaud
P
P
Perret
T
T
Grutter
M
M
Goeldner
D000345:Affinity Labels; D001665:Binding Sites; D003545:Cysteine / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005818:Genetic Engineering; D008024:Ligands; D016296:Mutagenesis; D018079:Receptors, GABA / Q000187:drug effects / Q000378:metabolism*; D016194:Receptors, N-Methyl-D-Aspartate / Q000187:drug effects / Q000378:metabolism
D000345:Affinity Labels; D008024:Ligands; D018079:Receptors, GABA; D016194:Receptors, N-Methyl-D-Aspartate; D003545:Cysteine