White P J, Ridout M
Horticulture Research International, Kent, United Kingdom.
J Membr Biol. 1995 Jan;143(1):37-49. doi: 10.1007/BF00232522.
The permeation of K+ and Na+ through the pore of a K+ channel from the plasma membrane of rye roots was studied in planar 1-palmitoyl-2-oleoyl phosphatidylethanolamine bilayers. The pore contains at least two ion-binding sites which can be occupied simultaneously. This was indicated by: (i) biphasic relationships with increasing cation concentration of both channel conductance at the zero-current (reversal) potential of the channel (Erev) and unitary-current at a specified voltage and (ii) a decline in Erev in the presence of equimolar Na+ (cis):K+ (trans) as the cation concentration was increased. To determine the spatial characteristics and energy profiles for K+ and Na+ permeation, unitary-current/voltage data for the channel were fitted to a three energy-barrier, two ion-binding site (3B2S) model. The model allowed for simultaneous occupancy of binding sites and ionic repulsion within the pore, as well as surface potential effects. Results suggested that energy peaks and energy wells (ion binding sites) were situated asymmetrically within the electrical distance of the pore, the trans energy-well being closer to the center of the pore than its cis counterpart; that the energy profile for K+ permeation differed significantly from that of Na+ in having a higher cis energy peak and a deeper cis energy well; that cations repelled each other within the pore and that vestibule surface charge was negligible. The model successfully simulated various aspects of K+ and Na+ permeation including: (i) the complexities in current rectification of a wide range of contrasting ionic conditions; (ii) the biphasic relationships with increasing cation concentration of both channel conductance at Erev and unitary-current at a specified voltage; (iii) the decline in Erev in equimolar Na+ (cis):K+ (trans) as cation concentrations were increased and (iv) the complex relationships between mole fraction and Erev at total cation concentrations of 100 and 300 mM.
在平面的1-棕榈酰-2-油酰磷脂酰乙醇胺双层膜中研究了钾离子(K⁺)和钠离子(Na⁺)通过黑麦根细胞质膜上钾离子通道孔的渗透情况。该通道孔至少包含两个可同时被占据的离子结合位点。这一点由以下现象表明:(i)在通道的零电流(反转)电位(Erev)下通道电导以及在特定电压下的单通道电流与阳离子浓度呈双相关系;(ii)当阳离子浓度增加时,在等摩尔的Na⁺(顺式):K⁺(反式)存在的情况下Erev下降。为了确定K⁺和Na⁺渗透的空间特征和能量分布,将通道的单通道电流/电压数据拟合到一个三能垒、双离子结合位点(3B2S)模型。该模型考虑了结合位点的同时占据、孔内的离子排斥以及表面电位效应。结果表明,能量峰和能量阱(离子结合位点)在孔的电距离内不对称分布,反式能量阱比顺式能量阱更靠近孔的中心;K⁺渗透的能量分布与Na⁺的显著不同,具有更高的顺式能量峰和更深的顺式能量阱;阳离子在孔内相互排斥,并且前庭表面电荷可忽略不计。该模型成功模拟了K⁺和Na⁺渗透的各个方面,包括:(i)在各种不同离子条件下电流整流的复杂性;(ii)在Erev下通道电导以及在特定电压下单通道电流与阳离子浓度呈双相关系;(iii)当阳离子浓度增加时,等摩尔的Na⁺(顺式):K⁺(反式)中Erev下降;(iv)在总阳离子浓度为100和300 mM时摩尔分数与Erev之间的复杂关系。