Korneeva N L, Jockusch B M
Cell Biology, Zoological Institute, Technical University of Braunschweig, Braunschweig, Germany.
Eur J Cell Biol. 1996 Dec;71(4):351-5.
In this study, we describe a simple light microscopic assay which allows to rapidly determine the ligand-induced organization of actin filaments into specific suprastructures, such as web-like arrangements or bundles. The validity of this assay is demonstrated by accompanying low shear (falling ball) viscometry. While the visually identified webs demonstrated viscosity values significantly higher than the F-actin control, the bundles were characterized by viscosities distinctly lower than that of the control. In addition, we show that at least in some cases, the type of actin suprastructure formed depends on the molar ratio between the ligand and actin filaments. The assay should be useful to assess the conditions under which a particular ligand leads to a specific actin filament organization, to determine quickly the biological activity of recombinant proteins or isolated actin-binding domains, and to define new F-actin ligands.
在本研究中，我们描述了一种简单的光学显微镜检测方法，该方法能够快速确定配体诱导的肌动蛋白丝组织形成特定的超结构，如网状排列或束状结构。伴随的低剪切（落球）粘度测定法证明了该检测方法的有效性。肉眼识别的网状结构的粘度值明显高于F-肌动蛋白对照，而束状结构的粘度则明显低于对照。此外，我们表明，至少在某些情况下，形成的肌动蛋白超结构类型取决于配体与肌动蛋白丝之间的摩尔比。该检测方法可用于评估特定配体导致特定肌动蛋白丝组织形成的条件，快速确定重组蛋白或分离的肌动蛋白结合结构域的生物学活性，以及定义新的F-肌动蛋白配体。
