Nakayasu T, Yokota E, Shimmen T
Department of Life Science, Faculty of Science, Himeji Institute of Technology, Hyogo, Japan.
Biochem Biophys Res Commun. 1998 Aug 10;249(1):61-5. doi: 10.1006/bbrc.1998.9088.
From lily pollen tubes, an actin-binding protein composed of 115-kDa polypeptide was purified sequentially by co-precipitation method with F-actin, hydroxylapatite column, gel filtration column and DE-52 ion exchange column chromatography. This component displayed a tendency to aggregate in solutions of low ionic strength, indicating a hydrophilic characteristic. Under physiological ionic conditions, this component bound to F-actin in an actin-concentration-dependent was saturable manner. Binding of this component to F-actin was independent of ATP and Ca(2+)-concentrations. Fluorescent microscopy revealed that F-actin labeled with rhodamine-phalloidin showed bundling in the presence of this component. Judging from the lack of antibody cross-reactivity, this component does not seem to be related to alpha-actinin of skeletal muscle and plant 135-kDa actin-bundling protein. Therefore, this component is the F-actin binding protein, which has not been identified thus far in plant cells.
从百合花粉管中，通过与F-肌动蛋白共沉淀法、羟基磷灰石柱、凝胶过滤柱和DE-52离子交换柱色谱法依次纯化出一种由115 kDa多肽组成的肌动蛋白结合蛋白。该组分在低离子强度溶液中有聚集倾向，表明具有亲水性。在生理离子条件下，该组分以肌动蛋白浓度依赖性且可饱和的方式与F-肌动蛋白结合。该组分与F-肌动蛋白的结合与ATP和Ca(2+)浓度无关。荧光显微镜显示，在该组分存在下，用罗丹明-鬼笔环肽标记的F-肌动蛋白呈束状。从缺乏抗体交叉反应性判断，该组分似乎与骨骼肌的α-辅肌动蛋白和植物135 kDa肌动蛋白束蛋白无关。因此，该组分是一种迄今在植物细胞中尚未鉴定出的F-肌动蛋白结合蛋白。
