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钠离子和钙离子通过非洲爪蟾卵母细胞中表达的小鼠ε2/ζ1 N-甲基-D-天冬氨酸受体通道的渗透特性。

Permeation properties of Na+ and Ca2+ ions through the mouse epsilon2/zeta1 NMDA receptor channel expressed in Xenopus oocytes.

作者信息

Iino M, Ciani S, Tsuzuki K, Ozawa S, Kidokoro Y

机构信息

Department of Physiology, Gunma University School of Medicine, 3-39-22 Showa-machi, Maebashi, 371, Japan.

出版信息

J Membr Biol. 1997 Jan 15;155(2):143-56. doi: 10.1007/s002329900166.

Abstract

Ion permeation properties of the mouse e2/zeta1 NMDA receptor channel expressed in Xenopus oocytes were studied using the outside-out patch-clamp technique. In symmetrical Na+ solutions, the single-channel I-V relations were almost linear at low electrolyte concentrations, but rectified inwardly for Na+ concentrations above 50 mm. In symmetrical Na+ solutions, the "zero-current conductance" increased with Na+ concentration and saturated according to a hyperbolic curve, the half-maximal saturating activity, KM(Na), being 14.2 mm and the maximal conductance, Gmax(Na), 53.9 pS. When Ca2+ was present with Na+ in the external solution, the single-channel current was lower than in pure Na+, although the reversal potential indicated a higher permeability for Ca2+ than for Na+. Using ion activities, PCa/PNa was found to be about 17. The I-V data were fitted with a model based on the Eyring's rate theory, assuming a one-ion pore with three energy barriers and two sites. The KM(Ca) and Gmax (Ca) were 76.5 microm and 21.2 pS, respectively. According to the estimated rate constants, KM for Ca2+ is mainly determined by the binding strength of a site located 80% away from the channel opening at the external membrane-solution interface, a position similar to that postulated previously for the Mg2+ blocking site.

摘要

利用外向膜片钳技术研究了非洲爪蟾卵母细胞中表达的小鼠e2/zeta1 N-甲基-D-天冬氨酸(NMDA)受体通道的离子通透特性。在对称的Na+溶液中,低电解质浓度下单通道电流-电压(I-V)关系几乎呈线性,但当Na+浓度高于50 mM时呈内向整流。在对称的Na+溶液中,“零电流电导”随Na+浓度增加,并根据双曲线曲线饱和,半最大饱和活性KM(Na)为14.2 mM,最大电导Gmax(Na)为53.9 pS。当外部溶液中Ca2+与Na+共存时,单通道电流低于纯Na+溶液中的电流,尽管反转电位表明Ca2+的通透性高于Na+。利用离子活度,发现PCa/PNa约为17。I-V数据用基于艾林速率理论的模型拟合,假设单离子孔有三个能垒和两个位点。KM(Ca)和Gmax (Ca)分别为76.5 μM和21.2 pS。根据估计的速率常数,Ca2+的KM主要由位于外膜-溶液界面距通道开口80%处的一个位点的结合强度决定,该位置与先前推测的Mg2+阻断位点的位置相似。

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