Bell J E, Miller C
Biophys J. 1984 Jan;45(1):279-87. doi: 10.1016/S0006-3495(84)84154-5.
Single-channel K+ currents through sarcoplasmic reticulum K+ channels were compared after reconstitution into planar bilayers formed from neutral or negatively charged phospholipids. In neutral bilayers, the channel conductance saturates with K+ concentration according to a rectangular hyperbola, with half-saturation at 40 mM K+, and maximum conductance of 220 pS. In negatively charged bilayers (70% phosphatidylserine/30% phosphatidylethanolamine), the conductance is, at a given K+ concentration, higher than in neutral bilayers. This effect of negative surface charge is increasingly pronounced at lower ionic strength. The maximum conductance at high K+ approaches 220 pS in negative bilayers, and the channel's ionic selectivity is unaffected by lipid charge. The divalent channel blocker " bisQ11 " causes discrete blocking events in both neutral and negatively charged bilayers; the apparent rate constant of blocking is sensitive to surface charge, while the unblocking rate is largely unaffected. Bilayers containing a positively charged phosphatidylcholine analogue led to K+ conductances lower than those seen in neutral bilayers. The results are consistent with a simple mechanism in which the local K+ concentration sensed by the channel's entryway is determined by both the bulk K+ concentration and the bulk lipid surface potential, as given by the Gouy-Chapman model of the electrified interface. To be described by this approach, the channel's entryway must be assumed to be located 1-2 nm away from the lipid surface, on both sides of the membrane.
将肌浆网钾通道重构到由中性或带负电荷的磷脂形成的平面双层膜中后,对单通道钾电流进行了比较。在中性双层膜中,通道电导根据矩形双曲线随钾离子浓度饱和,在40 mM钾离子时达到半饱和,最大电导为220 pS。在带负电荷的双层膜(70%磷脂酰丝氨酸/30%磷脂酰乙醇胺)中,在给定钾离子浓度下,电导高于中性双层膜。这种负表面电荷的影响在较低离子强度下越来越明显。在带负电荷的双层膜中,高钾离子浓度下的最大电导接近220 pS,且通道的离子选择性不受脂质电荷影响。二价通道阻滞剂“bisQ11”在中性和带负电荷的双层膜中都会引起离散的阻断事件;阻断的表观速率常数对表面电荷敏感,而解阻速率基本不受影响。含有带正电荷的磷脂酰胆碱类似物的双层膜导致钾离子电导低于中性双层膜中的电导。结果与一种简单机制一致,即通道入口处感知的局部钾离子浓度由本体钾离子浓度和本体脂质表面电位共同决定,如带电界面的古依-查普曼模型所给出的那样。为了用这种方法描述,必须假设通道入口位于膜两侧距脂质表面1 - 2 nm处。