Krasilnikov O V, Da Cruz J B, Yuldasheva L N, Varanda W A, Nogueira R A
Laboratory of Molecular Physiology, Institute of Physiology and Biophysics, 700095 Tashkent, Uzbekistan.
J Membr Biol. 1998 Jan 1;161(1):83-92. doi: 10.1007/s002329900316.
This paper describes a new approach to evaluate the inner structure (including a main constriction and its localization) of the water lumen of an ion channel. The method is based on the determination of channel filling by different nonelectrolyte molecules through each side of an ion channel. The method has two characteristic features that make its use attractive: (i) the possibility to ascertain the existence, localization and size of a narrow part inside an ion channel water lumen and (ii) the chances to determine the maximal size of both entrances of an ion channel and to obtain additional information about the geometry of its water lumen at the same time. Determinations were made on colicin Ia ion channels inserted into planar lipid bilayers. This channel was chosen because there is an apparent contradiction between its low single channel conductance and the large diameter of its water lumen. Our results show that the water lumen of the colicin Ia channel has a funnel-like structure with a small trans-entrance, with a diameter of about 1.0 nm, and a large cis-entrance, with a diameter of approximately 1.8 nm. A constriction with a diameter of approximately 0.7 nm is shown to be located close to the trans-entrance of the channel. The method can also be applied to patch clamp studies of single ion channels.
本文描述了一种评估离子通道水腔内部结构(包括一个主要收缩部位及其定位)的新方法。该方法基于通过离子通道的每一侧用不同的非电解质分子确定通道填充情况。该方法有两个使其具有吸引力的特点:(i)有可能确定离子通道水腔内狭窄部分的存在、定位和大小,以及(ii)有机会同时确定离子通道两个入口的最大尺寸,并获得有关其水腔几何形状的额外信息。对插入平面脂质双分子层的大肠杆菌素Ia离子通道进行了测定。选择这个通道是因为其低单通道电导与其水腔大直径之间存在明显矛盾。我们的结果表明,大肠杆菌素Ia通道的水腔具有漏斗状结构,有一个直径约为1.0纳米的小反式入口和一个直径约为1.8纳米的大顺式入口。已表明一个直径约为0.7纳米的收缩部位位于通道的反式入口附近。该方法也可应用于单离子通道的膜片钳研究。