Cingulin contains globular and coiled-coil domains and interacts with ZO-1, ZO-2, ZO-3, and myosin.
Cingulin contains globular and coiled-coil domains and interacts with ZO-1, ZO-2, ZO-3, and myosin.
复制标题
cingulin包含球形和盘绕螺旋域，并与ZO-1，ZO-2，ZO-3和肌球蛋白相互作用。
DOI：
10.1083/jcb.147.7.1569
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发表时间：
1999-12-27
影响因子：
7.8
通讯作者：
Citi, S
中科院分区：
文献类型：
--
作者：
Cordenonsi, M;D'Atri, F;Hammar, E;Parry, D A;Kendrick-Jones, J;Shore, D;Citi, S
我们的特点的序列和蛋白质相互作用的cingulin，一个M r 140-160 kD的磷蛋白定位于细胞质表面的上皮紧密连接（TJ）。全长非洲爪蟾扣带蛋白cDNA的衍生氨基酸序列显示球状头部（残基1-439）和尾部（1，326 - 1，368）结构域以及中央α-螺旋杆结构域（440- 1，325）。序列分析，电子显微镜，和下拉分析表明，扣带蛋白杆是负责形成卷曲螺旋平行二聚体，它可以通过分子间相互作用进一步聚集。上皮细胞、昆虫细胞和网织红细胞裂解物的拉下试验表明，扣带蛋白（1-378）的NH 2末端片段在体外与ZO-1（Kd ≤ 5 nM）、ZO-2、ZO-3、肌球蛋白和AF-6相互作用，但不与symplekin相互作用，COOH末端片段（377- 1，368）与肌球蛋白和ZO-3相互作用。ZO-1和ZO-2免疫沉淀物含有扣带蛋白，表明在体内相互作用。全长扣带蛋白，但不是NH 2-末端和COOH-末端片段，共定位与内源性扣带蛋白在转染的MDCK细胞，表明头和杆域内的序列是TJ本地化所需的。我们建议，扣带蛋白是一个功能上重要的组成部分，TJ，连接膜下斑块域TJ的肌动球蛋白细胞骨架。
We characterized the sequence and protein interactions of cingulin, an M r 140–160-kD phosphoprotein localized on the cytoplasmic surface of epithelial tight junctions (TJ). The derived amino acid sequence of a full-length Xenopus laevis cingulin cDNA shows globular head (residues 1–439) and tail (1,326–1,368) domains and a central α-helical rod domain (440–1,325). Sequence analysis, electron microscopy, and pull-down assays indicate that the cingulin rod is responsible for the formation of coiled-coil parallel dimers, which can further aggregate through intermolecular interactions. Pull-down assays from epithelial, insect cell, and reticulocyte lysates show that an NH2-terminal fragment of cingulin (1–378) interacts in vitro with ZO-1 (K d ∼5 nM), ZO-2, ZO-3, myosin, and AF-6, but not with symplekin, and a COOH-terminal fragment (377–1,368) interacts with myosin and ZO-3. ZO-1 and ZO-2 immunoprecipitates contain cingulin, suggesting in vivo interactions. Full-length cingulin, but not NH2-terminal and COOH-terminal fragments, colocalizes with endogenous cingulin in transfected MDCK cells, indicating that sequences within both head and rod domains are required for TJ localization. We propose that cingulin is a functionally important component of TJ, linking the submembrane plaque domain of TJ to the actomyosin cytoskeleton.