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兔骨骼肌肌浆网囊泡中两种类型氯离子通道的特性

Characteristics of two types of chloride channel in sarcoplasmic reticulum vesicles from rabbit skeletal muscle.

作者信息

Kourie J I, Laver D R, Junankar P R, Gage P W, Dulhunty A F

机构信息

John Curtin School of Medical Research, Australian National University, Canberra City, Australia.

出版信息

Biophys J. 1996 Jan;70(1):202-21. doi: 10.1016/S0006-3495(96)79564-4.

Abstract

A comparison is made of two types of chloride-selective channel in skeletal muscle sarcoplasmic reticulum (SR) vesicles incorporated into lipid bilayers. The I/V relationships of both channels, in 250/50 mM Cl- (cis/trans), were linear between -20 and +60 mV (cis potential,) reversed near Ecl and had slope conductances of approximately 250 pS for the big chloride (BCl) channel and approximately 70 pS for the novel, small chloride (SCl) channel. The protein composition of vesicles indicated that both channels originated from longitudinal SR and terminal cisternae. BCl and SCl channels responded differently to cis SO4(2-) (30-70 mM), 4,4'-diisothiocyanatostilbene 2,2'-disulfonic acid (8-80 microM) and to bilayer potential. The BCl channel open probability was high at all potentials, whereas SCl channels exhibited time-dependent activation and inactivation at negative potentials and deactivation at positive potentials. The duration and frequency of SCl channel openings were minimal at positive potentials and maximal at -40 mV, and were stationary during periods of activity. A substate analysis was performed using the Hidden Markov Model (S. H. Chung, J. B. Moore, L. Xia, L. S. Premkumar, and P. W. Gage, 1990, Phil. Trans. R. Soc. Lond. B., 329:265-285) and the algorithm EVPROC (evaluated here). SCl channels exhibited transitions between 5 and 7 conductance levels. BCl channels had 7-13 predominant levels plus many more short-lived substates. SCl channels have not been described in previous reports of Cl- channels in skeletal muscle SR.

摘要

对整合到脂质双分子层中的骨骼肌肌浆网(SR)囊泡中的两种氯离子选择性通道进行了比较。在250/50 mM Cl⁻(顺式/反式)条件下,两种通道的电流-电压(I/V)关系在-20至+60 mV(顺式电位)之间呈线性,在氯离子平衡电位(Ecl)附近反转,大氯离子(BCl)通道的斜率电导约为250 pS,新型小氯离子(SCl)通道的斜率电导约为70 pS。囊泡的蛋白质组成表明,两种通道均起源于纵向SR和终池。BCl和SCl通道对顺式SO₄²⁻(30 - 70 mM)、4,4'-二异硫氰基芪-2,2'-二磺酸(8 - 80 μM)以及双分子层电位的反应不同。BCl通道在所有电位下开放概率都很高,而SCl通道在负电位下表现出时间依赖性的激活和失活,在正电位下表现为去激活。SCl通道开放的持续时间和频率在正电位下最小,在-40 mV时最大,并且在活动期间保持稳定。使用隐马尔可夫模型(S. H. Chung、J. B. Moore、L. Xia、L. S. Premkumar和P. W. Gage,1990年,《英国皇家学会哲学学报B》,329:265 - 285)和算法EVPROC(在此评估)进行了亚状态分析。SCl通道表现出5至7个电导水平之间的转换。BCl通道有7 - 13个主要水平以及更多寿命较短的亚状态。在先前关于骨骼肌SR中氯离子通道的报道中尚未描述过SCl通道。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f1d/1224921/601f625ffab2/biophysj00052-0205-a.jpg

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