Identification of a novel intracellular interaction domain essential for Bves function.
Identification of a novel intracellular interaction domain essential for Bves function.
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DOI：
10.1371/journal.pone.0002261
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发表时间：
2008-05-21
期刊：
影响因子：
3.7
通讯作者：
Bader DM
中科院分区：
文献类型：
--
作者：
Kawaguchi M;Hager HA;Wada A;Koyama T;Chang MS;Bader DM
虽然血管心外膜物质（Bves）赋予粘附特性，但调节这种活性的分子机制尚不清楚。没有预测的功能基序，在这个高度保守的膜蛋白，除跨膜结构域，已被确定。在这里，我们报告的第一次，Bves通过细胞内的相互作用域，这是必不可少的细胞间粘附活性与自身相互作用。谷胱甘肽-S-转移酶（GST）下拉和SPOTs分析将该结构域映射到细胞内C-末端的氨基酸268-274。定点突变显示赖氨酸272和273对于同源二聚化和细胞粘附是必需的。用野生型Bves转染的人角膜细胞将蛋白质运输到细胞表面，组装连接复合物并形成上皮片。相比之下，在这些位置突变的表达Bves的细胞不形成连续的上皮片或维持连接蛋白，如ZO-1和E-钙粘蛋白在膜上。还观察到跨上皮电阻的急剧降低，表明紧密连接的功能丧失。重要的是，突变的Bves在上皮细胞中的表达促进了细胞从上皮表型向间充质表型的转化。这项研究是第一个证明Bves内任何结构域维持上皮表型和功能的基本性质。
While Blood vessel epicardial substance (Bves) confers adhesive properties, the molecular mechanism of regulating this activity is unknown. No predicted functional motifs in this highly conserved integral membrane protein, other than the transmembrane domain, have been identified. Here, we report for the first time that Bves interacts with itself through an intracellular interaction domain that is essential for its intercellular adhesion activity. Glutathion-S-transferase (GST) pull-down and SPOTs analyses mapped this domain to amino acids 268-274 in the intracellular C-terminus. Site-directed mutagenesis revealed that lysines 272 and 273 are essential for homodimerization and cell adhesion. Human corneal cells transfected with wild-type Bves trafficked the protein to the cell surface, assembled junction complexes and formed epithelial sheets. In contrast, cells expressing Bves mutated at these positions did not form continuous epithelial sheets or maintain junctional proteins such as ZO-1 and E-cadherin at the membrane. A dramatic reduction in transepithelial electrical resistance was also observed indicating a functional loss of tight junctions. Importantly, expression of mutated Bves in epithelial cells promoted the transformation of cells from an epithelial to a mesenchymal phenotype. This study is the first to demonstrate the essential nature of any domain within Bves for maintenance of epithelial phenotype and function.
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影响因子：
4.8
作者：
Knight, RF;Bader, DM;Backstrom, JR
通讯作者：
Backstrom, JR
影响因子：
2.5
作者：
Breher, SS;Mavridou, E;Brand, T
通讯作者：
Brand, T
DOI：
10.1083/jcb.148.1.189
发表时间：
2000-01-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Thoreson MA;Anastasiadis PZ;Daniel JM;Ireton RC;Wheelock MJ;Johnson KR;Hummingbird DK;Reynolds AB
通讯作者：
Reynolds AB
影响因子：
2.7
作者：
Andrée, B;Hillemann, T;Brand, T
通讯作者：
Brand, T
DOI：
10.1073/pnas.0506095103
发表时间：
2006-01-17
影响因子：
11.1
作者：
Ripley, AN;Osler, ME;Bader, D
通讯作者：
Bader, D