The role of conserved aspartate and serine residues in ligand binding and in function of the 5-HT1A receptor: a site-directed mutation study.
Wild-type and mutant serotonin 1A receptors were transiently expressed in COS-7 cells using the infection-transfection variant of the vaccinia virus/T7 polymerase vector system. The amino acid substitutions in the transmembrane regions, Asp82-->Asn82, Asp116-->Asn116, and Ser198-->Ala198 all resulted in a decrease in affinity for 5-HT by 60-100-fold, without affecting the affinity for the antagonist, pindolol. The binding of agonist to the additional mutant, Thr199-->Ala199, was too weak to be measured, 5-HT induced GTPase activities for all receptors studied. These findings indicate that the residues mutated play an important role in the binding of the agonist and less critical roles in the binding of the antagonist pindolol.
Ho
B Y
BY
Division of Biology, California Institute of Technology, Pasadena 91125.
Karschin
A
A
Branchek
T
T
Davidson
N
N
Lester
H A
HA
D001224:Aspartic Acid / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D020558:GTP Phosphohydrolases / Q000378:metabolism; D008024:Ligands; D016297:Mutagenesis, Site-Directed; D011985:Receptors, Serotonin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*
D008024:Ligands; D011985:Receptors, Serotonin; D001224:Aspartic Acid; D012694:Serine; D020558:GTP Phosphohydrolases