Zaner K S, Hartwig J H
Massachusetts General Hospital, Department of Medicine, Harvard Medical School, Boston 02114.
J Biol Chem. 1988 Apr 5;263(10):4532-6.
To address the claim that filaments polymerized from highly purified (gel-filtered) F-actin acquire the elastic properties of a solid attributable to chemical cross-linking, we measured the rheologic spectrum of the dynamic storage modulus, G', and loss modulus, G'' from 5 x 10(-4) to 0.5 Hz for gel-filtered actin alone and in the presence of the actin shortening protein, gelsolin. We confirmed that gel-filtered filamentous actin is a highly elastic material as evidenced by a relatively frequency-independent G', which is consistent with either topologically constrained filaments or a chemically cross-linked gel. Introduction of gel-filtered actin oligomers, however, caused the behavior of gel-filtered actin to become more frequency-dependent and almost identical to that of non-gel-filtered actin, suggesting that the effect of gel filtration on the mechanical behavior of actin is topologic. This conclusion is further supported by the finding that shortening of the actin filaments by the addition of gelsolin at molar ratios to actin of from 1:8000 to 1:500 causes a gradual decrease in elasticity and increase in the amount of flow.
为了回应关于从高度纯化(凝胶过滤)的F-肌动蛋白聚合而成的细丝获得了归因于化学交联的固体弹性特性这一说法,我们测量了单独的凝胶过滤肌动蛋白以及在存在肌动蛋白缩短蛋白凝溶胶蛋白的情况下,从5×10⁻⁴到0.5赫兹的动态储能模量G'和损耗模量G''的流变学频谱。我们证实,凝胶过滤的丝状肌动蛋白是一种高弹性材料,这表现为相对频率无关的G',这与拓扑受限的细丝或化学交联的凝胶是一致的。然而,引入凝胶过滤的肌动蛋白寡聚体导致凝胶过滤肌动蛋白的行为变得更依赖频率,并且几乎与未凝胶过滤的肌动蛋白相同,这表明凝胶过滤对肌动蛋白力学行为的影响是拓扑学上的。通过添加凝溶胶蛋白使肌动蛋白丝缩短,当凝溶胶蛋白与肌动蛋白的摩尔比从1:8000到1:500时,弹性逐渐降低且流动量增加,这一发现进一步支持了这一结论。