Figl A, Labarca C, Davidson N, Lester H A, Cohen B N
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena 91125, USA.
J Gen Physiol. 1996 Mar;107(3):369-79. doi: 10.1085/jgp.107.3.369.
We have studied the voltage-jump relaxation currents for a series of neuronal nicotinic acetylcholine receptors resulting from the coexpression of wild-type and chimeric beta 4/beta 2 subunits with alpha 3 subunits in Xenopus oocytes. With acetylcholine as the agonist, the wild-type alpha 3 beta 4 receptors displayed five- to eightfold slower voltage-jump relaxations than did the wild-type alpha 3 beta 2 receptors. In both cases, the relaxations could best be described by two exponential components of approximately equal amplitudes over a wide range of [ACh]'s. Relaxation rate constants increased with [ACh] and saturated at 20- to 30-fold lower concentrations for the alpha 3 beta 2 receptor than for the alpha 3 beta 4 receptor, as observed previously for the peak steady state conductance. Furthermore, the chimeric beta 4/beta 2 subunits showed a transition in the concentration dependence of the rate constants in the region between residues 94 and 109, analogous to our previous observation with steady state conductances. However, our experiments with a series of beta-subunit chimeras did not localize residues that govern the absolute value of the kinetic parameters. Hill coefficients for the relaxations also differed from those previously measured for steady state responses. The data reinforce previous conclusions that the region between residues 94 and 109 on the beta subunit plays a role in binding agonist but also show that other regions of the receptor control gating kinetics subsequent to the binding step.
我们研究了一系列神经元烟碱型乙酰胆碱受体的电压阶跃松弛电流,这些受体是通过在非洲爪蟾卵母细胞中共同表达野生型和嵌合的β4/β2亚基与α3亚基产生的。以乙酰胆碱作为激动剂时,野生型α3β4受体的电压阶跃松弛比野生型α3β2受体慢5至8倍。在这两种情况下,在很宽的乙酰胆碱浓度范围内,松弛最好用两个幅度大致相等的指数成分来描述。松弛速率常数随乙酰胆碱浓度增加而增加,并且α3β2受体在比α3β4受体低20至30倍的浓度时达到饱和,这与之前观察到的峰值稳态电导情况相同。此外,嵌合的β4/β2亚基在残基94和109之间的区域显示出速率常数浓度依赖性的转变,这与我们之前对稳态电导的观察类似。然而,我们对一系列β亚基嵌合体的实验并未定位到控制动力学参数绝对值的残基。松弛的希尔系数也与之前测量的稳态反应的希尔系数不同。这些数据强化了之前的结论,即β亚基上残基94和109之间的区域在结合激动剂中起作用,但也表明受体的其他区域在结合步骤之后控制门控动力学。