Shelley Christopher, Colquhoun David
Department of Pharmacology, University College London, London WC1E 6BT, UK.
J Physiol. 2005 Apr 15;564(Pt 2):377-96. doi: 10.1113/jphysiol.2004.081497. Epub 2005 Feb 24.
A mutation in the epsilon subunit of the human nicotinic acetylcholine receptor (epsilonL78P) is known to cause a congenital slow channel myasthenic syndrome. We have investigated the changes in receptor function that result in the mutant receptor producing prolonged endplate currents, and consequent muscle damage. The rate constants for channel gating and for the binding and dissociation of acetylcholine were investigated by analysis of single ion channel recordings. A conventional mechanism with two non-equivalent binding sites, and variations upon this mechanism, were fitted to data using a maximum likelihood method that uses the Hawkes-Jalali-Colquhoun (HJC) treatment of missed brief events. The mutant receptor produced prolonged activations, bursts of openings that cause a slow decay of simulated synaptic currents. The main reason for the longer bursts of openings seen with mutant receptor was a decrease in the rate of ACh dissociation from diliganded receptors, though the lifetime of individual openings was somewhat increased too. As well as producing long bursts, the mutant receptor also produced many very short openings, though these carry little current. The burst structure for the mutant receptor at low ACh concentration is unusual in that most long bursts appear to start in a very brief monoliganded open state that then usually binds another ACh molecule to produce a long diliganded activation. The first opening is so short that it will usually be missed (together with the shut time that follows it), so the true burst length is likely to be underestimated.
已知人类烟碱型乙酰胆碱受体的ε亚基发生突变(εL78P)会导致先天性慢通道重症肌无力综合征。我们研究了受体功能的变化,这些变化导致突变型受体产生延长的终板电流,进而导致肌肉损伤。通过对单离子通道记录的分析,研究了通道门控以及乙酰胆碱结合和解离的速率常数。使用一种最大似然方法,将具有两个不等价结合位点的传统机制及其变体与数据进行拟合,该方法采用了霍克斯 - 贾拉利 - 科尔库洪(HJC)对错过的短暂事件的处理方法。突变型受体产生了延长的激活,即开放爆发,导致模拟突触电流缓慢衰减。突变型受体出现较长开放爆发的主要原因是乙酰胆碱从双配体受体解离的速率降低,不过单个开放的持续时间也略有增加。除了产生长爆发外,突变型受体还产生了许多非常短暂的开放,尽管这些开放携带的电流很小。低乙酰胆碱浓度下突变型受体的爆发结构不同寻常,因为大多数长爆发似乎始于一个非常短暂的单配体开放状态,然后通常会结合另一个乙酰胆碱分子以产生长时间的双配体激活。第一次开放非常短暂,以至于通常会被错过(连同随后的关闭时间),因此实际的爆发长度可能被低估。