Site-directed mutagenesis studies on the Drosophila octopamine/tyramine receptor.
The cloned Drosophila octopamine/tyramine receptor can be coupled to second messenger pathways in an agonist-specific fashion by the endogenously occurring biogenic amines, octopamine and tyramine, when expressed in Chinese hamster ovary cells. We have mutated to alanine a range of receptor amino acids that could potentially form hydrogen bonds with the beta-hydroxyl group of octopamine based on homologies with alpha- and beta-adrenergic receptor subtypes. After stable expression of the mutant receptors in CHO cells we have compared the ability of octopamine and tyramine to displace [(3)H]yohimbine binding to membrane fractions from the mutant cell lines with their ability to modulate adenylyl cyclase activity in intact cells. The results suggest that none of the mutated amino acids residues, at least in isolation, are likely to be involved in interactions with the beta-hydroxyl group of the octopamine side chain. It is possible that amino acids not mutated in the present study are somehow involved in this interaction. Alternatively, it is also possible that the beta-hydroxyl group of the octopamine side chain is capable of interacting with more than one of the amino acids mutated in the present study.
Chatwin
Heather M
HM
The Laboratory of Receptor Signalling, The Babraham Institute, Babraham, Cambridge CB2 4AT, UK.
Rudling
Jane E
JE
Patel
Daksha
D
Reale
Vincenzina
V
Evans
Peter D
PD
D019943:Amino Acid Substitution; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D004330:Drosophila / Q000235:genetics*; D006860:Hydrogen Bonding; D016297:Mutagenesis, Site-Directed; D009655:Octopamine / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D017982:Receptors, Biogenic Amine / Q000235:genetics*; D011994:Recombinant Proteins / Q000378:metabolism; D014162:Transfection; D015016:Yohimbine / Q000378:metabolism
D009838:Oligodeoxyribonucleotides; D017982:Receptors, Biogenic Amine; D011994:Recombinant Proteins; C027604:norsynephrine receptor; C066175:tyramine receptor; D009655:Octopamine; D015016:Yohimbine