Differential binding properties of [H-3]dextrorphan and [H-3]MK-801 in heterologously expressed NMDA receptors

• Type of Material: Article
• Authors: K.T. LePage, University of Georgia; J.E. Ishmael, Oregon State University; C.M. Low, National University of Singapore; Stephen Traynelis, Emory University; T.F. Murray, University of Georgia
• Language: English
• Date: 2005-07-01
• Publisher: PERGAMON-ELSEVIER SCIENCE LTD
• Publication Version: Author Accepted Manuscript (After Peer Review)
• Copyright Statement: © 2005 Elsevier Ltd.
• Final Published Version (URL): http://dx.doi.org/10.1016/j.neuropharm.2005.01.029
• Title of Journal or Parent Work: Neuropharmacology
• Volume: 49
• Issue: 1
• Start Page: 1
• End Page: 16
• Grant/Funding Information: Supported by National Institute on Drug Abuse grant (DA07218) to T.F.M., an F32 National Research Service Award (DA05728) to J.E.I., and NINDS grant (NS36654) to S.F.T.
• Keywords: Science & Technology; GLUTAMATE; NMDA receptor; Neurosciences & Neurology; EXTRACELLULAR VESTIBULE; SUBSTITUTED CYSTEINES; PHENCYCLIDINE; RAT-BRAIN; D-ASPARTATE RECEPTOR; MK-801; Pharmacology & Pharmacy; PHARMACOLOGY & PHARMACY; NEUROSCIENCES; CHANNEL BLOCKERS; MOLECULAR DETERMINANTS; Neurosciences; Life Sciences & Biomedicine; glutamate; mutagenesis; STRUCTURAL DETERMINANTS; dextrorphan; binding; SITE
• Research Categories: Biology, Neuroscience; Health Sciences, Pharmacology

Abstract

The N-methyl-D-aspartate receptor (NMDAR) antagonists: MK-801, phencyclidine and ketamine are open-channel blockers with limited clinical value due to psychotomimetic effects. Similarly, the psychotomimetic effects of the dextrorotatory opioids, dextromethorphan and its metabolite dextrorphan, derive from their NMDAR antagonist actions. Differences in the use dependency of blockade, however, suggest that the binding sites for MK-801 and dextrorphan are distinct. In the absence of exogenous glutamate and glycine, the rate of association of [3H]MK-801 with wild-type NR1-1a/NR2A receptors was considerably slower than that for [3H]dextrorphan. Glutamate individually, and in the presence of the co-agonist glycine, had substantial effects on the specific binding of [3H]MK-801, while the binding of [3H]dextrorphan was not affected. Mutation of residues N616 and A627 in the NR1 subunit had a profound effect on [3H]MK-801 binding affinity, while that of [3H]dextrorphan was unaltered. In contrast, NR1 residues, W611 and N812, were critical for specific binding of [ 3H]dextrorphan to NR1-1a/NR2A complexes with no corresponding influence on that of [3H]MK-801. Thus, [3H]dextrorphan and [3H]MK-801 have distinct molecular determinants for high-affinity binding. The ability of [3H]dextrorphan to bind to a closed channel, moreover, indicates that its recognition site is shallower in the ion channel domain than that of MK-801 and may be associated with the extracellular vestibule of the NMDAR. © 2005 Elsevier Ltd. All rights reserved.