A unique binding epitope for salvinorin A, a non-nitrogenous kappa opioid receptor agonist.
Salvinorin A is a potent kappa opioid receptor (KOP) agonist with unique structural and pharmacological properties. This non-nitrogenous ligand lacks nearly all the structural features commonly associated with opioid ligand binding and selectivity. This study explores the structural basis to salvinorin A binding and selectivity using a combination of chimeric and single-point mutant opioid receptors. The experiments were designed based on previous models of salvinorin A that locate the ligand within a pocket formed by transmembrane (TM) II, VI, and VII. More traditional sites of opioid recognition were also explored, including the highly conserved aspartate in TM III (D138) and the KOP selectivity site E297, to determine the role, if any, that these residues play in binding and selectivity. The results indicate that salvinorin A recognizes a cluster of residues in TM II and VII, including Q115, Y119, Y312, Y313, and Y320. Based on the position of these residues within the receptor, and prior study on salvinorin A, a model is proposed that aligns the ligand vertically, between TM II and VII. In this orientation, the ligand spans residues that are spaced one to two turns down the face of the helices within the receptor cavity. The ligand is also in close proximity to EL-2 which, based on chimeric data, is proposed to play an indirect role in salvinorin A binding and selectivity.
Kane
Brian E
BE
Department of Medicinal Chemistry and Center for Drug Design, University of Minnesota, MN 55455, USA.
Nieto
Marcelo J
MJ
McCurdy
Christopher R
CR
Ferguson
David M
DM
D000818:Animals; D001665:Binding Sites; D002460:Cell Line; D002478:Cells, Cultured; D004224:Diterpenes / Q000378:metabolism*; D045785:Diterpenes, Clerodane; D018604:Epitope Mapping; D000939:Epitopes / Q000378:metabolism*; D006801:Humans; D051379:Mice; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D051381:Rats; D017465:Receptors, Opioid, delta / Q000235:genetics / Q000378:metabolism; D017473:Receptors, Opioid, kappa / Q000819:agonists* / Q000235:genetics / Q000378:metabolism*; D017450:Receptors, Opioid, mu / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D027544:Salvia / Q000737:chemistry* / Q000378:metabolism
D004224:Diterpenes; D045785:Diterpenes, Clerodane; D000939:Epitopes; D017465:Receptors, Opioid, delta; D017473:Receptors, Opioid, kappa; D017450:Receptors, Opioid, mu; D011993:Recombinant Fusion Proteins; C090499:salvinorin A