Colocalization of F-actin and 34-kilodalton actin bundling protein in Dictyostelium amoebae and cultured fibroblasts.
Colocalization of F-actin and 34-kilodalton actin bundling protein in Dictyostelium amoebae and cultured fibroblasts.
复制标题
F-肌动蛋白和 34 千道尔顿肌动蛋白捆绑蛋白在阿米巴盘基网柄菌和培养的成纤维细胞中的共定位。
DOI：
10.1002/cm.970090303
复制
发表时间：
1988
影响因子：
--
通讯作者：
Pardee,JD
中科院分区：
文献类型：
--
作者：
Johns,JA;Brock,AM;Pardee,JD
从盘状网柄藻分离的Ca+2-敏感肌动蛋白结合蛋白，30，000-D蛋白（Fechheimer和Taylor：J. Biol. Chem.259：4514-4520，1984;）最近已定位于底物粘附阿米巴的丝状伪足中（Fechheimer：J. Cell Biol.104：1539-1551，1987）。我们已经确定，这种蛋白质具有34，000道尔顿的Mr，并且在底物附着的Dictyosteliumamoeacin和培养的成纤维细胞中与肌动蛋白丝严格共定位。3 T3成纤维细胞，以及正常和病毒转化的大鼠肾成纤维细胞（NRK）含有一种34 kD的蛋白质，该蛋白质与网柄藻捆绑蛋白的抗体特异性交叉反应。哺乳动物34 kD蛋白与应力纤维中的F肌动蛋白和基质粘附成纤维细胞中的皮质细胞骨架共定位。在基质粘附的植物性网骨藻中，F-肌动蛋白和34-kD蛋白质集中在细胞皮质的显示丝状伪足和膜脊的区域。暴露于化学引诱物叶酸后形成的多个丝状伪足强烈染色34 kD蛋白，这意味着在丝状伪足形成期间参与肌动蛋白束的组装。伪足皮层也含有高浓度的捆绑蛋白，但伪足内部没有。与植物性Dictyosteoblasts相反，F-actin和34-kD蛋白在发育周期中进展的细胞中没有共定位。在子实体中，通过免疫荧光显微镜仅在前孢子细胞中检测到34-kD蛋白，而F-肌动蛋白出现在柄细胞和孢子中。
The Ca+2‐sensitive actin‐binding protein isolated fromDictyostelium discoideum, 30,000‐D protein (Fechheimer and Taylor:J. Biol. Chem.259:4514–4520, 1984;) has recently been localized in filipodia of substrate‐adhered amoebae (Fechheimer:J. Cell Biol.104:1539–1551, 1987). We have determined that this protein has a Mrof 34,000 daltons and is strictly colocalized with actin filaments in both substrate‐attachedDictyosteliumamoebae and cultured fibroblasts. 3T3 fibroblasts, as well as normal and virally transformed rat kidney fibroblasts (NRK) contain a 34‐kilodalton (kD) protein that cross‐reacts specifically with antibody to theDictyosteliumbundling protein. Mammalian 34‐kD protein is colocalized with F‐actin in stress fibers and the cortical cytoskeleton in substratadhered fibroblasts. In substrate‐adhered vegetativeDictyostelium, F‐actin and 34‐kD protein are concentrated in regions of the cell cortex exhibiting filipodia and membrane ridges. Multiple filipodia formed after exposure to the chemoattractant folic acid stain intensely for 34‐kD protein, implying participation in the assembly of actin bundles during filipod formation. The cortex of pseudopodia also contained high concentrations of bundling protein, but pseudopod interiors did not. In contrast to vegetativeDictyostelium, F‐actin and 34‐kD protein were not colocalized in cells that had progressed through the development cycle. In fruiting bodies, 34‐kD protein was detected by immunofluorescence microscopy only in prespore cells, while F‐actin appeared in stalk cells and spores.