Tuszynski J A, Brown J A, Sept D
Department of Physics, University of Alberta, Edmonton, Alberta T6G 2J1 Canada.
J Biol Phys. 2003 Dec;29(4):401-28. doi: 10.1023/A:1027318920964.
One of the most important issues of molecular biophysics is the complex and multifunctional behavior of the cell's cytoskeleton. Interiors of living cells are structurally organized by the cytoskeleton networks of filamentous protein polymers: microtubules, actin and intermediate filaments with motor proteins providing force and directionality needed for transport processes. Microtubules (MT's) take active part in material transport within the cell, constitute the most rigid elements of the cell and hence found many uses in cell motility (e.g. flagella andcilia). At present there is, however, no quantitatively predictable explanation of how these important phenomena are orchestrated at a molecular level. Moreover, microtubules have been demonstrated to self-organize leading to pattern formation. We discuss here several models which attempt to shed light on the assembly of microtubules and their interactions with motor proteins. Subsequently, an overview of actin filaments and their properties isgiven with particular emphasis on actin assembly processes. The lengths of actin filaments have been reported that were formed by spontaneous polymerization of highly purified actin monomers after labeling with rhodamine-phalloidin. The length distributions are exponential with a mean of about 7 μm. This length is independent of the initial concentration of actin monomer, an observation inconsistent with a simple nucleation-elongation mechanism. However, with the addition of physically reasonable rates of filament annealing and fragmenting, a nucleation-elongation mechanism can reproduce the observed average length of filaments in two types of experiments: (1) filaments formed from a wide range of highly purified actin monomer concentrations, and (2) filaments formed from 24 mM actin over a range of CapZ concentrations. In the final part of the paper we briefly review the stochastic models used to describe the motion of motor proteins on protein filaments. The vast majority of these models are based on ratchet potentials with the presence of thermal noise and forcing due to ATP binding and a subsequent hydrolysis. Many outstanding questions remain to be quantitatively addressed on a molecular level in order to explain the structure-to-function relationship for the key elements of the cytoskeleton discussed in this review.
分子生物物理学最重要的问题之一是细胞细胞骨架的复杂和多功能行为。活细胞内部由丝状蛋白质聚合物的细胞骨架网络进行结构组织:微管、肌动蛋白和中间丝,以及运动蛋白提供运输过程所需的力和方向性。微管(MT)积极参与细胞内的物质运输,构成细胞中最坚硬的元素,因此在细胞运动(如鞭毛和纤毛)中有许多用途。然而,目前对于这些重要现象如何在分子水平上协调进行,还没有定量可预测的解释。此外,微管已被证明能够自我组织并导致图案形成。我们在此讨论几种试图阐明微管组装及其与运动蛋白相互作用的模型。随后,对肌动蛋白丝及其特性进行了概述,特别强调了肌动蛋白组装过程。在用罗丹明 - 鬼笔环肽标记后,报道了由高度纯化的肌动蛋白单体自发聚合形成的肌动蛋白丝的长度。长度分布呈指数分布,平均值约为7μm。这个长度与肌动蛋白单体的初始浓度无关,这一观察结果与简单的成核 - 伸长机制不一致。然而,通过添加物理上合理的丝退火和断裂速率,成核 - 伸长机制可以在两种类型的实验中重现观察到的丝的平均长度:(1)由广泛的高度纯化的肌动蛋白单体浓度形成的丝,以及(2)在一系列CapZ浓度下由24 mM肌动蛋白形成的丝。在本文的最后部分,我们简要回顾了用于描述运动蛋白在蛋白质丝上运动的随机模型。这些模型中的绝大多数基于棘轮势,存在热噪声以及由于ATP结合和随后的水解产生的驱动力。为了解释本综述中讨论的细胞骨架关键元件的结构 - 功能关系,在分子水平上仍有许多突出问题有待定量解决。