Pan X X, Ware B R
Department of Chemistry, Syracuse University, New York 13244-1200.
Biophys J. 1988 Jan;53(1):11-6. doi: 10.1016/S0006-3495(88)83060-1.
The ability of Li+ to promote the assembly of actin has been compared with the more common cations used in actin assembly assays, K+, Mg2+, and Ca2+. The principal assay of actin assembly utilized was fluorescence photobleaching recovery (FPR), from which it is possible to determine the fraction of actin protomers incorporated into filaments and the average diffusion coefficients of the filaments. In addition, critical concentrations of actin over a range of concentrations of all of these cations have been determined using an assay that involves sonication and dilution of assembled actin filaments containing trace amounts of pyrene-labeled actin. The results demonstrate that Li+ is a more potent promoter of actin assembly than is K+. The more rapid assembly of actin in the presence of Li+ is attributable to an increased rate of filament elongation. Filaments assembled in equivalent concentrations of Li+ or K+ have the same diffusion coefficients, and thus presumably the same average lengths. The critical concentration of actin is about three times less in the presence of Li+ than in the presence of an equal concentration of K+. Cytochalasin D accelerates the rate of Li+-promoted actin assembly and reduces slightly the total fraction of actin assembly. However, cytochalasin D causes less shortening of filaments in the presence of Li+ than in the presence of K+ or Mg2+. By the criteria of assembly kinetics and critical concentration, Li+ is much less potent as a promoter of actin assembly than either Mg2+ or Ca2+. These results are discussed in terms of the role of electrostatic forces in the actin assembly mechanism and in terms of possible relationships to therapeutic and toxicity mechanisms for Li+.
已将Li+促进肌动蛋白组装的能力与肌动蛋白组装测定中常用的其他阳离子(K+、Mg2+和Ca2+)进行了比较。所使用的主要肌动蛋白组装测定方法是荧光光漂白恢复(FPR),通过该方法可以确定掺入细丝中的肌动蛋白原聚体的比例以及细丝的平均扩散系数。此外,使用一种涉及对含有微量芘标记肌动蛋白的组装肌动蛋白细丝进行超声处理和稀释的测定方法,确定了在所有这些阳离子的一系列浓度下肌动蛋白的临界浓度。结果表明,Li+比K+更有效地促进肌动蛋白组装。在Li+存在下肌动蛋白组装更快是由于细丝伸长率增加。在等效浓度的Li+或K+中组装的细丝具有相同的扩散系数,因此推测平均长度相同。在Li+存在下肌动蛋白的临界浓度比在相同浓度的K+存在下约低三倍。细胞松弛素D加速了Li+促进的肌动蛋白组装速率,并略微降低了肌动蛋白组装的总比例。然而,在Li+存在下,细胞松弛素D导致细丝缩短的程度比在K+或Mg2+存在下小。根据组装动力学和临界浓度的标准,Li+作为肌动蛋白组装促进剂的效力远低于Mg2+或Ca2+。将根据静电力在肌动蛋白组装机制中的作用以及与Li+的治疗和毒性机制的可能关系来讨论这些结果。