Kroemker M, Rüdiger A H, Jockusch B M, Rüdiger M
Department of Cell Biology, Technical University Braunschweig, Germany.
FEBS Lett. 1994 Dec 5;355(3):259-62. doi: 10.1016/0014-5793(94)01216-4.
Using blot overlay techniques we have investigated the interaction of vinculin with alpha-actinin. We show that an alpha-actinin binding site is located in the 90 kDa vinculin head and confirm a vinculin binding site in the C-terminal rod of alpha-actinin, as recently reported by McGregor et al. [(1994) Biochem. J. 310, 225-233]. The isolated vinculin head binds much more strongly to alpha-actinin than intact vinculin. Using a proteolytic 81 kDa head fragment, we show that vinculin residues 1-107 are required for alpha-actinin binding. Antibodies directed against vinculin residues 808-850 inhibit the vinculin-alpha-actinin binding, suggesting that this sequence is directly involved in, or topographically related to, the alpha-actinin binding site.
我们运用印迹覆盖技术研究了纽蛋白与α-辅肌动蛋白的相互作用。我们发现α-辅肌动蛋白结合位点位于90 kDa的纽蛋白头部，并证实了α-辅肌动蛋白C末端杆状区域存在纽蛋白结合位点，这与麦格雷戈等人最近的报道一致[（1994年）《生物化学杂志》310卷，225 - 233页]。分离出的纽蛋白头部与α-辅肌动蛋白的结合比完整的纽蛋白更强。使用一个81 kDa的蛋白水解头部片段，我们发现纽蛋白1 - 107位残基是与α-辅肌动蛋白结合所必需的。针对纽蛋白808 - 850位残基的抗体抑制了纽蛋白与α-辅肌动蛋白的结合，这表明该序列直接参与α-辅肌动蛋白结合位点或与该结合位点在拓扑结构上相关。
