Journal
BIOSENSORS & BIOELECTRONICS
Volume 15, Issue 3-4, Pages 173-181Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/S0956-5663(00)00058-0
Keywords
agonist selectivity; epsilon 1/zeta 1 N-methyl-D-aspartate receptor channel; N-methyl-D-aspartate; L-glutamate; (2S, 3R, 4S)-2-(carboxycyclopropyl)glycine; tip-dip method
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A new method for evaluating chemical selectivity of agonists to activate the N-methyl-D-aspartate (NMDA) receptor was presented by using typical agonists NMDA, L-glutamate and (2S, 3R, 4S)-2-(carboxycyclopropyl)glycine (L-CCG-IV) and the mouse epsilon 1/zeta 1 NMDA receptor incorporated in bilayer lipid membranes (BLMs) as an illustrative example. The method was based on the magnitude of an agonist-induced integrated single-channel current corresponding to the number of total ions passed through the open channel. The very magnitudes of the integrated single-channel currents were compared with the different BLMs as a new measure of agonist selectivity. The epsilon 1/zeta 1 NMDA receptor was partially purified from Chinese hamster ovary (CHO) cells expressing the epsilon 1/zeta 1 NMDA receptor and incorporated in BLMs formed by the tip-dip method. The agonist-induced integrated single-channel currents were obtained at 50 mu M agonist concentration, where the integrated current for NMDA was shown to reach its saturated value. The obtained integrated currents were found to be (4.5 +/- 0.55) x 10(-13) C/s for NMDA, (5.8 +/- 0.72) x 10(-13) C/s for L-glutamate and (6.6 +/- 0.61) x 10(-13) C/s for L-CCG-IV, respectively. These results suggest that the agonist selectivity in terms of the total ion flux through the single epsilon 1/zeta 1 NMDA receptor is in the order of L-CCG-IV approximate to L-glutamate > NMDA. (C) 2000 Elsevier Science S.A. All rights reserved.
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