Mutation analysis of the short cytoplasmic domain of the cell-cell adhesion molecule CEACAM1 identifies residues that orchestrate actin binding and lumen formation
Mutation analysis of the short cytoplasmic domain of the cell-cell adhesion molecule CEACAM1 identifies residues that orchestrate actin binding and lumen formation
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DOI：
10.1074/jbc.m610903200
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发表时间：
2007-02-23
影响因子：
4.8
通讯作者：
Shively, John E.
中科院分区：
文献类型：
--
作者：
Chen, Charng-Jui;Kirshner, Julia;Shively, John E.
在乳腺形态发生的三维模型中，CEACAM1-4S（癌胚抗原细胞粘附分子1，具有4个外结构域和一个短的12-14个氨基酸的细胞质结构域）在乳腺上皮细胞生长过程中通过凋亡和细胞骨架重组机制介导管腔形成。我们通过定量酵母双杂交、BIAcore、NMR HSQC和STD以及共聚焦分析表明，苯丙氨酸（Phe(454)）和赖氨酸（Lys(456)）是与肌动蛋白相互作用协调细胞骨架重组的关键残基。基于维生素d结合蛋白的CEACAM1膜模型预测了肌动蛋白亚结构域3上Phe 454的相互作用，该模型得到了细胞质结构域肽抑制肌动蛋白与维生素d结合蛋白结合的支持。我们还发现，在三维培养中，转染ceacam1的细胞中，残基Thr（457）和/或Ser（419）被磷酸化，这些残基（T457A/S459A）或F454A的突变分析阻断了管腔的形成。这些研究表明，短胞质结构域膜受体可以直接介导大量的细胞内信号传导。
CEACAM1-4S (carcinoembryonic antigen cell adhesion molecule 1, with 4 ectodomains and a short, 12-14 amino acid cytoplasmic domain) mediates lumen formation via an apoptotic and cytoskeletal reorganization mechanism when mammary epithelial cells are grown in a three-dimensional model of mammary morphogenesis. We show by quantitative yeast two-hybrid, BIAcore, NMR HSQC and STD, and confocal analyses that amino acids phenylalanine (Phe(454)) and lysine (Lys(456)) are key residues that interact with actin orchestrating the cytoskeletal reorganization. A CEACAM1 membrane model based on vitamin D-binding protein that predicts an interaction of Phe 454 at subdomain 3 of actin was supported by inhibition of binding of actin to vitamin D-binding protein by the cytoplasmic domain peptide. We also show that residues Thr(457) and/or Ser(419) are phosphorylated in CEACAM1-transfected cells grown in three-dimensional culture and that mutation analysis of these residues (T457A/S459A) or F454A blocks lumen formation. These studies demonstrate that a short cytoplasmic domain membrane receptor can directly mediate substantial intracellular signaling.