An electrophilic affinity ligand based on (+)-MK801 distinguishes PCP site 1 from PCP site 2 |
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Authors: | Hyacinth C Akunne J A Monn Andrew Thurkauf Arthur E Jacobson Kenner C Rice Joannes T M Linders Q Jiang F Porreca Richard B Rothman |
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Institution: | (1) Clinical Psychopharmacology Section, NIDA/NIH Addiction Research Center, P.O. Box 5180, 21224 Baltimore, MD;(2) Laboratory of Medicinal Chemistry, NIDDK, 20892 Bethesda, MD;(3) Department of Pharmacology, University of Arizona, 85724 Tucson, Arizona |
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Abstract: | The electrophilic affinity ligand, (+)-3-isothiocyanato-5-methyl-10,11-dihydro-5H-dibenzoa,d] cyclohepten-5,10-imine hydrochloride {(+)-MK801-NCS} was characterized for its ability to acylate phencyclidine (PCP) and sigma binding sites in vivo. Initial studies, conducted with mouse brain membranes, characterized the binding sites labeled by 3H]1-1-(2-thienyl)cyclohexyl]piperidine (3H]TCP). The Kd values of 3H]TCP for PCP site 1 (MK801-sensitive) and PCP site 2 (MK801-insensitive) were 12 nM and 68 nM, with Bmax values of 1442 and 734 fmol/mg protein, respectively. Mice were sacrificed 18–24 hours following intracerebroventricular administration of the acylator. The administration of (+)-MK801-NCS increased 3H]TCP binding to site 2, but not to site. 1. Although (+)-MK801-NCS decreased 3H](+)-5-methyl-10,11-dihydro-5H-dibenzoa,d; cbcyclohepten-5,10-imine maleate (3H](+)-MK801) binding to site 1, it had no effect on 3H]TCP binding to site 1. Viewed collectively with other published data, these data support the hypothesis that PCP sites 1 and 2 are distinct binding sites, and that 3H]TCP and 3H](+)-MK801 label different domains of the PCP binding site associated with the NMDA receptor.Abbreviations ((+)-MK801)
(+)-5-methyl-10,11-dihydro-5H-dibenzoa,d]cyclohepten-5,10-imine
- ((+)-MK801-NCS)
(+)-3-isothiocyanato-5-methyl-10,11-dihydro-5H-dibenzoa,d] cyclohepten-5,10-imine hydrochloride
- (PCP)
1-(1-phencyclohexyl)piperidine
- (TCP)
1-{1-(2-thienyl)cyclohexyl}piperidine
- (DTG)
(2-(tllyl)guanidine
- (metaphit)
(1-(1-(3-isothiocyanatophenyl)-cyclohexyl)piperidine)
- (NMDA)
N-methyl-D-aspartate
- (HEPPSO)
(N-2-hydroxyethyl]piperazine-N-2-hydroxypropanesulfoni c acid]
- ((+)-MK801-NCS)
(+)-5-methyl(3-isothiocyanatophenyl)-10,11-dihydro-5H-dibenzoa,d]cyclohepten-5,10-imine
- (NMDA)
N-methyl-D-aspartate
Address reprint requests to Dr. Rothman, Phone (410)550-1487.FAX 410-550-2997 |
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Keywords: | N-methyl-D-aspartate (+)-MK801 affinity ligands PCP 1 3-di(2-tolyl)guanidine sigma receptors |
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