Klüsener B, Boheim G, Liss H, Engelberth J, Weiler E W
Department of Plant Physiology, Ruhr-University, Bochum, Germany.
EMBO J. 1995 Jun 15;14(12):2708-14. doi: 10.1002/j.1460-2075.1995.tb07271.x.
The lipid bilayer technique was adapted to the functional reconstitution of ion channels from the endoplasmic reticulum of a higher plant. This was obtained at high purity from touch-sensitive tendrils of Bryonia dioica. In this preparation, a calcium-selective strongly rectifying channel is prevailing whose single-channel properties have been characterized. The single-channel conductance is 29 pS in 50 mM CaCl2. The Ca2+: K+ selectivity was determined to be approximately 6.6. The channel is voltage-gated and, more importantly, the gating voltage is strongly shifted towards more negative voltages when a transmembrane Ca2+ gradient is applied. Thus, at physiological voltages across the endoplasmic reticulum membrane, the channel's open probability will be governed largely by the chemical potential gradient of Ca2+, generated by the Ca(2+)-ATPase in that same membrane. The calcium release channel described here is effectively blocked by Gd3+ which also completely suppresses a tendril's reaction to touch, suggesting that this channel could be a key element of calcium signaling in higher plant mechanotransduction. Its molecular characteristics and inhibitor data show it to be the first known member of a hitherto unrecognized class of calcium channels.
脂质双分子层技术被应用于高等植物内质网离子通道的功能重建。该离子通道是从白泻根的触敏卷须中高纯度提取得到的。在此制备过程中,一种钙选择性强整流通道占主导地位,其单通道特性已得到表征。在50 mM氯化钙中,单通道电导为29 pS。测定的Ca2+:K+选择性约为6.6。该通道是电压门控的,更重要的是,当施加跨膜Ca2+梯度时,门控电压会强烈地向更负的电压偏移。因此,在内质网膜两侧的生理电压下,通道的开放概率将主要由同一膜中Ca(2+)-ATP酶产生的Ca2+化学势梯度决定。这里描述的钙释放通道被Gd3+有效阻断,Gd3+也完全抑制了卷须对触摸的反应,这表明该通道可能是高等植物机械转导中钙信号传导的关键元件。其分子特征和抑制剂数据表明它是迄今未被识别的一类钙通道中的首个已知成员。