Buss F, Temm-Grove C, Henning S, Jockusch B M
Cell Biology Unit, University of Bielefeld, Federal Republic of Germany.
Cell Motil Cytoskeleton. 1992;22(1):51-61. doi: 10.1002/cm.970220106.
We have used polyclonal and monoclonal antibodies raised against calf thymus profilin to localize the corresponding protein in translocating, spreading, and stationary rat fibroblasts. Immunofluorescence of whole cells and immunogold labeling on ventral membranes of lysis-squirted cells showed that profilin was markedly enriched in the highly dynamic lamellipodia or pseudopodial lobes. Within these regions, a significant fraction was colocalized with dynamic actin filaments organized in actin ribs, cortical filaments, or stress fiber-like bundles, and little profilin was found in membrane areas appearing free of actin. In contrast, stress fibers of stationary cells as well as actin arcs and ring-like bundles of spreading and migrating cells showed very little label. These results are discussed in context with the proposed role of profilin in regional membrane dynamics typical for fibroblasts and are compared to previous data (Hartwig et al.: J. Cell Biol. 109:1571-1579, 1989) on profilin distribution in platelets and granulocytes.
我们使用了针对小牛胸腺肌动蛋白结合蛋白产生的多克隆抗体和单克隆抗体,以在迁移、铺展和静止的大鼠成纤维细胞中定位相应的蛋白质。全细胞免疫荧光以及裂解喷射细胞腹侧膜上的免疫金标记显示,肌动蛋白结合蛋白在高度动态的片状伪足或伪足叶中显著富集。在这些区域内,很大一部分与排列成肌动蛋白肋、皮质细丝或应力纤维样束状的动态肌动蛋白丝共定位,而在没有肌动蛋白的膜区域几乎没有发现肌动蛋白结合蛋白。相比之下,静止细胞的应力纤维以及铺展和迁移细胞的肌动蛋白弧和环状束显示出很少的标记。这些结果结合肌动蛋白结合蛋白在成纤维细胞典型的区域膜动力学中所提出的作用进行了讨论,并与之前关于肌动蛋白结合蛋白在血小板和粒细胞中分布的数据(Hartwig等人:《细胞生物学杂志》109:1571 - 1579,1989年)进行了比较。