Supervillin (p205): A novel membrane-associated, F-actin-binding protein in the villin/gelsolin superfamily.
Supervillin (p205): A novel membrane-associated, F-actin-binding protein in the villin/gelsolin superfamily.
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DOI：
10.1083/jcb.139.5.1255
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发表时间：
1997-12-01
影响因子：
7.8
通讯作者：
Luna, EJ
中科院分区：
文献类型：
--
作者：
Pestonjamasp, KN;Pope, RK;Wulfkuhle, JD;Luna, EJ
肌动蛋白结合膜蛋白参与粘附相互作用和运动过程。我们在这里报告的纯化和初步表征p205，205 kD的蛋白质从牛中性粒细胞质膜，结合到两侧的肌动蛋白丝的印迹覆盖。p205是一种紧密结合的外周膜蛋白，在蔗糖梯度中与内源性肌动蛋白共沉淀并免疫沉淀。从SDS-PAGE纯化的p205中获得氨基酸序列，并用于产生抗肽抗体、免疫定位数据和cDNA序列信息。p205在MDBK细胞中的细胞内定位是细胞密度和粘附状态的函数。在亚融合细胞中，p205被发现在点状斑点沿着质膜和细胞质和细胞核中;在贴壁细胞中，p205集中与E-钙粘蛋白在网站的横向细胞-细胞接触。在EGTA介导的细胞解离后，p205与E-钙粘蛋白和F-肌动蛋白一起内化，作为粘附连接“环”的组分。在稍后的时间，在细胞质点中观察到p205。在中性粒细胞和悬浮生长的HeLa细胞，缺乏adherens连接，p205的高丰度，进一步表明，这种蛋白质可能在细胞生长，粘附和运动过程中发挥多种作用。p205 cDNA的分子克隆揭示了一个二分结构。COOH末端与绒毛蛋白和凝溶胶蛋白具有惊人的相似性，特别是在已知结合F-肌动蛋白的区域。NH 2末端是新的，但包含四个潜在的核靶向信号。由于p205是目前已知的绒毛蛋白/凝溶胶蛋白超家族中最大的成员，我们建议将其命名为“超绒毛蛋白”。我们认为，超绒毛蛋白可能参与肌动蛋白丝组装在adherens交界处，它可能在其他细胞隔室发挥额外的作用。
Actin-binding membrane proteins are involved in both adhesive interactions and motile processes. We report here the purification and initial characterization of p205, a 205-kD protein from bovine neutrophil plasma membranes that binds to the sides of actin filaments in blot overlays. p205 is a tightly bound peripheral membrane protein that cosediments with endogenous actin in sucrose gradients and immunoprecipitates. Amino acid sequences were obtained from SDS-PAGE–purified p205 and used to generate antipeptide antibodies, immunolocalization data, and cDNA sequence information. The intracellular localization of p205 in MDBK cells is a function of cell density and adherence state. In subconfluent cells, p205 is found in punctate spots along the plasma membrane and in the cytoplasm and nucleus; in adherent cells, p205 concentrates with E-cadherin at sites of lateral cell–cell contact. Upon EGTA-mediated cell dissociation, p205 is internalized with E-cadherin and F-actin as a component of adherens junctions “rings.” At later times, p205 is observed in cytoplasmic punctae. The high abundance of p205 in neutrophils and suspension-grown HeLa cells, which lack adherens junctions, further suggests that this protein may play multiple roles during cell growth, adhesion, and motility. Molecular cloning of p205 cDNA reveals a bipartite structure. The COOH terminus exhibits a striking similarity to villin and gelsolin, particularly in regions known to bind F-actin. The NH2 terminus is novel, but contains four potential nuclear targeting signals. Because p205 is now the largest known member of the villin/gelsolin superfamily, we propose the name, “supervillin.” We suggest that supervillin may be involved in actin filament assembly at adherens junctions and that it may play additional roles in other cellular compartments.
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