Bowlby M R, Levitan I B
Biochemistry Department, Brandeis University, Waltham, MA 02254, USA.
Neuropharmacology. 1996;35(7):867-75. doi: 10.1016/0028-3908(96)00090-1.
We have examined, using patch recording, the modulation by ATP gamma S of the cloned Drosophila slopoke calcium-dependent potassium channel (dSlo) expressed in Xenopus oocytes. There is a large variation in the gating kinetics, open probability, and conductance level of the channel in this expression system, which complicates the analysis of modulatory events. Addition of ATP gamma S to the intracellular face of the patch does not consistently alter the overall open probability of dSlo, but it does increase the frequency of appearance of an exceptionally long-lived closed state of the channel. This modulation is not blocked by an inhibitor of several serine/threonine protein kinases, nor by mutation of a serine residue that is a target for phosphorylation by protein kinase A. Thus, ATP gamma S may alter dSlo kinetic properties by some mechanism other than serine/threonine phosphorylation.
我们利用膜片钳记录技术,研究了ATPγS对非洲爪蟾卵母细胞中表达的克隆果蝇慢poke钙依赖性钾通道(dSlo)的调制作用。在这个表达系统中,该通道的门控动力学、开放概率和电导水平存在很大差异,这使得对调制事件的分析变得复杂。将ATPγS添加到膜片的胞内面并不能始终如一地改变dSlo的总体开放概率,但它确实增加了通道出现异常长寿关闭状态的频率。这种调制作用既不被几种丝氨酸/苏氨酸蛋白激酶的抑制剂所阻断,也不被作为蛋白激酶A磷酸化靶点的丝氨酸残基突变所阻断。因此,ATPγS可能通过丝氨酸/苏氨酸磷酸化以外的某种机制改变dSlo的动力学特性。