Oda T, Makino K, Yamashita I, Namba K, Maéda Y
International Institute for Advanced Research, Matsushita Electric Industrial Co., Kyoto 619-0237, Japan.
Biophys J. 1998 Dec;75(6):2672-81. doi: 10.1016/S0006-3495(98)77712-4.
We examined factors that affect the filament orientation in F-actin sols to prepare highly well-oriented liquid crystalline sols suitable for x-ray fiber diffraction structure analysis. Filamentous particles such as F-actin spontaneously align with one another when concentrated above a certain threshold concentration. This alignment is attributed to the excluded volume effect of the particles. In trying to improve the orientation of F-actin sols, we focused on the excluded volume to see how it affects the alignment. The achievable orientation was sensitive to the ionic strength of the solvent; the filaments were better oriented at lower ionic strengths, where the effective diameter of the filament is relatively large. Sols of longer filaments were better oriented than those of shorter filaments at the same concentration, but the best achievable orientation was limited, probably because of the filament flexibility. The best strategy for making well-oriented F-actin sols is therefore to concentrate F-actin filaments of relatively short length (<1 micrometer) by slow centrifugation in a low-ionic-strength solvent (<30 mM).
我们研究了影响F-肌动蛋白溶胶中细丝取向的因素,以制备适用于X射线纤维衍射结构分析的高度取向良好的液晶溶胶。当丝状颗粒如F-肌动蛋白浓缩到高于一定阈值浓度时,它们会自发地相互排列。这种排列归因于颗粒的排阻体积效应。在试图改善F-肌动蛋白溶胶的取向时,我们关注排阻体积,以了解其如何影响排列。可实现的取向对溶剂的离子强度敏感;在较低离子强度下,细丝取向更好,此时细丝的有效直径相对较大。在相同浓度下,较长细丝的溶胶比较短细丝的溶胶取向更好,但可实现的最佳取向是有限的,这可能是由于细丝的柔韧性。因此,制备取向良好的F-肌动蛋白溶胶的最佳策略是在低离子强度溶剂(<30 mM)中通过低速离心浓缩相对较短长度(<1微米)的F-肌动蛋白细丝。