title: Hydrophobic interactions of n-alkyl diamines with the N-methyl-D-aspartate receptor: voltage-dependent and -independent blocking sites.
abstract: We examined the block of N-methyl-D-aspartate (NMDA) receptors by n-alkyl (straight chain) diamines and related monoamines and triamines using whole-cell voltage clamp recording of NMDA receptor currents in cultured rat hippocampal neurons and [3H] dizocilpine binding to rat forebrain homogenates. At -60 mV, the diamines (carbon chain lengths 3-12) produced a concentration-dependent inhibition of NMDA receptor current (IC50 values, 6128-7.3 microM). For diamines of carbon chain lengths greater than 6, the inhibition was partially, but not completely, relieved by depolarization, indicating that the block occurs at distinct voltage-dependent and voltage-independent sites. The block produced by short-chain diamines (carbon chain lengths 3-6) was completely relieved by depolarization, indicating little or no interaction with the voltage-independent site. In comparison with the corresponding diamines, homologous monoamines exhibited very low potency, whereas homologous triamines were of equal or lower potency. For long-chain diamines, inhibitory potency at both the voltage-dependent and voltage-independent sites was correlated with carbon chain length (binding energy increasing 600-700 cal/mol-CH2), suggesting that binding to each of the sites is stabilized by a hydrophobic interaction. Affinities for the voltage-dependent blocking site (transformed to 0 mV) and for the voltage-independent blocking site were similar. These values were also similar to the inhibitory potencies of the diamines in the [3H]dizocilpine binding assay. Analysis of the voltage-dependence of block at the voltage-dependent site yielded z delta values for diamines of intermediate length (carbon chain lengths 7-9) that decreased with increasing length from 0.91 to 0.63 [approaching the z delta values of monovalent blockers (approximately 0.54) and one-half of the z delta values of shorter diamines (approximately 1.1)], suggesting that the intermediate length diamines block in a linear, extended chain conformation with one of the charges having incomplete access to a deep binding site. Longer chain diamines (carbon chain lengths 10 and 12) exhibited larger z delta values (0.78 and 0.98, respectively), presumably because enhanced conformational flexibility permitted a folded-over conformation. From the interchange distances of the intermediate length diamines in their lowest energy conformation, we estimated that the total voltage drop within the NMDA receptor channel occurs over a distance of approximately 20 A. The putative polyamine facilitatory site antagonist diethylenetriamine inhibited NMDA-induced currents at the voltage-dependent site (IC50, 654 microM; -60 mV).(ABSTRACT TRUNCATED AT 400 WORDS)
authors.affiliation: Neuronal Excitability Section, National Institute of Neurological Diseases and Stroke, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: S
authors.initials: S
authors.lastname: Subramaniam
authors.forename: S D
authors.initials: SD
authors.lastname: Donevan
authors.forename: M A
authors.initials: MA
authors.lastname: Rogawski
mesh_terms: D000818:Animals; D001665:Binding Sites; D002478:Cells, Cultured; D003959:Diamines / Q000737:chemistry / Q000494:pharmacology*; D016291:Dizocilpine Maleate / Q000037:antagonists & inhibitors / Q000378:metabolism; D004563:Electrochemistry; D011073:Polyamines / Q000378:metabolism / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D016194:Receptors, N-Methyl-D-Aspartate / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D003959:Diamines; D011073:Polyamines; D016194:Receptors, N-Methyl-D-Aspartate; C005391:diethylenetriamine; D016291:Dizocilpine Maleate