Direct inhibition of the actomyosin motility by local anesthetics in vitro.
Direct inhibition of the actomyosin motility by local anesthetics in vitro.
复制标题
体外局部麻醉剂直接抑制肌动球蛋白运动。
DOI：
10.1016/s0006-3495(96)79466-3
复制
发表时间：
1996
影响因子：
3.4
通讯作者：
T. Yanagida
中科院分区：
文献类型：
--
作者：
Y. Tsuda;T. Mashimo;I. Yoshiya;K. Kaseda;Y. Harada;T. Yanagida
使用最近开发的体外运动试验，我们已经证明，局部麻醉剂直接抑制肌球蛋白为基础的运动的单个肌动蛋白丝在一个可逆的剂量依赖性的方式。这是第一次报道的帐户的作用，局部麻醉剂对纯化的蛋白质在分子水平上。在这项研究中，两个叔胺局部麻醉药，利多卡因和丁卡因，被使用。局部麻醉剂对肌动球蛋白滑动运动的抑制作用是pH依赖性的;麻醉剂在较高的pH值下更有效，并且这种反应伴随着不带电形式的麻醉剂的比例增加。QX-314是利多卡因的永久性带电衍生物，对肌动球蛋白滑动运动没有影响。这些结果表明，不带电形式的局部麻醉药是主要负责的肌动球蛋白滑动运动的抑制。局麻药抑制滑动运动，但几乎不干扰肌动蛋白丝的表面上的肌球蛋白的结合或肌动球蛋白ATP酶活性在低离子强度。为了表征在麻醉剂存在下的肌动球蛋白相互作用，我们测量了肌动球蛋白复合物的结合力和断裂力。在低离子强度下，利多卡因不影响肌动蛋白丝与肌球蛋白表面的结合。使用光镊测量的断裂力为每微米肌动蛋白丝约1.5 pN，这远小于僵硬力和等距力。结合力和断裂力随着离子强度的增加而大大降低，表明剩余的相互作用本质上是离子的。结果表明，肌动球蛋白的结合和ATP酶主要由离子相互作用控制，这是几乎不受麻醉剂的影响;而力的产生需要疏水相互作用，这起着重要作用的强结合，被麻醉剂阻断，除了离子相互作用。
Using a recently developed in vitro motility assay, we have demonstrated that local anesthetics directly inhibit myosin-based movement of single actin filaments in a reversible dose-dependent manner. This is the first reported account of the actions of local anesthetics on purified proteins at the molecular level. In this study, two tertiary amine local anesthetics, lidocaine and tetracaine, were used. The inhibitory action of the local anesthetics on actomyosin sliding movement was pH dependent; the anesthetics were more potent at higher pH values, and this reaction was accompanied by an increased proportion of the uncharged form of the anesthetics. QX-314, a permanently charged derivative of lidocaine, had no effect on actomyosin sliding movement. These results indicate that the uncharged form of local anesthetics is predominantly responsible for the inhibition of actomyosin sliding movement. The local anesthetics inhibited sliding movement but hardly interfered with the binding of actin filaments to myosin on the surface or with actomyosin ATPase activity at low ionic strength. To characterize the actomyosin interaction in the presence of anesthetics, we measured the binding and breaking force of the actomyosin complex. The binding of actin filaments to myosin on the surface was not affected by lidocaine at low ionic strength. The breaking force, measured using optical tweezers, was approximately 1.5 pN per micron of an actin filament, which was much smaller than in rigor and isometric force. The binding and breaking force greatly decreased with increasing ionic strength, indicating that the remaining interaction is ionic in nature. The result suggests that the binding and ATPase of actomyosin are governed predominantly by ionic interaction, which is hardly affected by anesthetics; whereas the force generation requires hydrophobic interaction, which plays a major part of the strong binding and is blocked by anesthetics, in addition to the ionic interaction.
影响因子：
--
作者：
HOMSHER, E;WANG, F;SELLERS, JR
通讯作者：
SELLERS, JR
DOI：
10.1073/pnas.83.17.6272
发表时间：
1986-09-01
影响因子：
11.1
作者：
KRON, SJ;SPUDICH, JA
通讯作者：
SPUDICH, JA
影响因子：
56.9
作者：
RAGSDALE, DS;MCPHEE, JC;CATTERALL, WA
通讯作者：
CATTERALL, WA