Renault A, Lenne P F, Zakri C, Aradian A, Vénien-Bryan C, Amblard F
Laboratoire de Spectrométrie Physique, Centre National de la Recherche Scientifique, UMR-5588, BP87, 38402 St Martin d'Heres, France.
Biophys J. 1999 Mar;76(3):1580-90. doi: 10.1016/S0006-3495(99)77317-0.
Living cells contain a very large amount of membrane surface area, which potentially influences the direction, the kinetics, and the localization of biochemical reactions. This paper quantitatively evaluates the possibility that a lipid monolayer can adsorb actin from a nonpolymerizing solution, induce its polymerization, and form a 2D network of individual actin filaments, in conditions that forbid bulk polymerization. G- and F-actin solutions were studied beneath saturated Langmuir monolayers containing phosphatidylcholine (PC, neutral) and stearylamine (SA, a positively charged surfactant) at PC:SA = 3:1 molar ratio. Ellipsometry, tensiometry, shear elastic measurements, electron microscopy, and dark-field light microscopy were used to characterize the adsorption kinetics and the interfacial polymerization of actin. In all cases studied, actin follows a monoexponential reaction-limited adsorption with similar time constants (approximately 10(3) s). At a longer time scale the shear elasticity of the monomeric actin adsorbate increases only in the presence of lipids, to a 2D shear elastic modulus of mu approximately 30 mN/m, indicating the formation of a structure coupled to the monolayer. Electron microscopy shows the formation of a 2D network of actin filaments at the PC:SA surface, and several arguments strongly suggest that this network is indeed causing the observed elasticity. Adsorption of F-actin to PC:SA leads more quickly to a slightly more rigid interface with a modulus of mu approximately 50 mN/m.
活细胞含有大量的膜表面积,这可能会影响生化反应的方向、动力学和定位。本文定量评估了在禁止本体聚合的条件下,脂质单层能否从非聚合溶液中吸附肌动蛋白、诱导其聚合并形成单个肌动蛋白丝的二维网络的可能性。在摩尔比为PC:SA = 3:1的含有磷脂酰胆碱(PC,中性)和硬脂胺(SA,一种带正电荷的表面活性剂)的饱和朗缪尔单层膜下研究了G-肌动蛋白和F-肌动蛋白溶液。使用椭偏仪、张力测定法、剪切弹性测量、电子显微镜和暗场光学显微镜来表征肌动蛋白的吸附动力学和界面聚合。在所研究的所有情况下,肌动蛋白遵循具有相似时间常数(约10³秒)的单指数反应限制吸附。在更长的时间尺度上,单体肌动蛋白吸附物的剪切弹性仅在脂质存在下增加,达到约30 mN/m的二维剪切弹性模量,表明形成了与单层膜耦合的结构。电子显微镜显示在PC:SA表面形成了肌动蛋白丝的二维网络,并有力地表明这个网络确实导致了观察到的弹性。F-肌动蛋白吸附到PC:SA上会更快地导致界面稍微更坚硬,模量约为50 mN/m。