Ruhnau K, Wegner A
Institute of Physiological Chemistry, Ruhr-University Bochum, FRG.
FEBS Lett. 1988 Feb 8;228(1):105-8. doi: 10.1016/0014-5793(88)80595-7.
The interaction of vinculin with actin filaments was investigated by methods which exclude interference by contaminating proteins which may occur in vinculin preparations. Vinculin which was blotted from SDS-polyacrylamide gels onto nitrocellulose, was stained specifically by fluorescently labeled polymeric actin (100 mM KCl, 2 mM MgCl2). Vinculin which was purified from alpha-actinin and an actin polymerization-inhibiting protein (HA1), was found to be cosedimented with polymeric actin. Maximally one vinculin molecule was cosedimented per one hundred actin filament subunits. Half maximal binding of vinculin was observed at about 0.25 microM free vinculin. Vinculin could be replaced from actin by the addition of tropomyosin.
通过排除纽蛋白制剂中可能存在的污染蛋白干扰的方法，研究了纽蛋白与肌动蛋白丝的相互作用。从SDS-聚丙烯酰胺凝胶转移到硝酸纤维素膜上的纽蛋白，被荧光标记的聚合肌动蛋白（100 mM KCl，2 mM MgCl2）特异性染色。从α-辅肌动蛋白和一种肌动蛋白聚合抑制蛋白（HA1）中纯化得到的纽蛋白，被发现与聚合肌动蛋白共沉降。每一百个肌动蛋白丝亚基中最多有一个纽蛋白分子共沉降。在游离纽蛋白浓度约为0.25 microM时观察到纽蛋白的半最大结合。通过添加原肌球蛋白，纽蛋白可从肌动蛋白上被取代。
