Helfer Emmanuèle, Panine Pierre, Carlier Marie-France, Davidson Patrick
Dynamique du Cytosquelette, Laboratoire d'Enzymologie et Biochimie Structurales, UPR 9063 Centre National de la Recherche Scientifique, Gif-sur-Yvette, France.
Biophys J. 2005 Jul;89(1):543-53. doi: 10.1529/biophysj.104.050245. Epub 2005 Apr 29.
F-actin gels of increasing concentrations (25-300 microM) display in vitro a progressive onset of birefringence due to orientational ordering of actin filaments. At F-actin concentrations <100 microM, this birefringence can be erased and restored at will by sonication and gentle flow, respectively. Hence, the orientational ordering does not result from a thermodynamic transition to a nematic phase but instead is due to mechanical stresses stored in the gels. In contrast, at F-actin concentrations > or =100 microM, gels display spontaneous birefringence recovery, at rest, which is the sign of true nematic ordering, in good agreement with statistical physics models of the isotropic/nematic transition. Well-aligned samples of F-actin gels could be produced and their small-angle x-ray scattering patterns are quite anisotropic. These patterns show no sign of filament positional short-range order and could be modeled by averaging the form factor with the Maier-Saupe nematic distribution function. The derived nematic order parameter S of the gels ranged from S = 0.7 at 300 microM to S = 0.4 at 25 microM. Both birefringence and small-angle x-ray scattering data indicate that, even in absence of cross-linking proteins, spontaneous cooperative alignment of actin filaments may arise in motile regions of living cells where F-actin concentrations can reach values of a few 100 microM.
浓度不断增加(25 - 300微摩尔)的F - 肌动蛋白凝胶在体外由于肌动蛋白丝的取向有序化而呈现出双折射的逐渐出现。在F - 肌动蛋白浓度小于100微摩尔时,这种双折射可以分别通过超声处理和缓慢流动随意消除和恢复。因此,取向有序化并非源于向向列相的热力学转变,而是由于凝胶中储存的机械应力。相比之下,在F - 肌动蛋白浓度大于或等于100微摩尔时,凝胶在静止时会呈现出自发的双折射恢复,这是真正向列有序化的标志,与各向同性/向列转变的统计物理模型高度吻合。可以制备出排列良好的F - 肌动蛋白凝胶样品,其小角X射线散射图案具有相当的各向异性。这些图案没有显示出细丝位置短程有序的迹象,并且可以通过将形状因子与迈尔 - 索佩向列分布函数进行平均来建模。所推导的凝胶向列序参数S的范围从300微摩尔时的S = 0.7到25微摩尔时的S = 0.4。双折射和小角X射线散射数据均表明,即使在没有交联蛋白的情况下,肌动蛋白丝的自发协同排列也可能出现在活细胞的运动区域,其中F - 肌动蛋白浓度可以达到几百微摩尔的值。