Dissociation of Madin-Darby canine kidney epithelial cells by the monoclonal antibody anti-arc-1: mechanistic aspects and identification of the antigen as a component related to uvomorulin.
Dissociation of Madin-Darby canine kidney epithelial cells by the monoclonal antibody anti-arc-1: mechanistic aspects and identification of the antigen as a component related to uvomorulin.
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DOI：
10.1083/jcb.101.4.1307
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发表时间：
1985-10
期刊：
影响因子：
--
通讯作者：
Imhof BA
中科院分区：
文献类型：
--
作者：
Behrens J;Birchmeier W;Goodman SL;Imhof BA
已有研究表明，抗Arc-1的单抗可在体外分离狗肾上皮细胞并改变其形态(Imhof，B.A.，H.P.Vollmers，S.L.Goodman，and W.Birchmeier，1983，Cell，35：667-675)。在这篇文章中，我们证明了抗Arc-1抗体识别MDCK细胞上的葡萄胎蛋白样分子，即在有钙离子存在的情况下，它从细胞表面的胰酶消化中免疫沉淀一段84kD的蛋白质片段(与抗葡萄膜蛋白抗血清一样)。此外，抗葡萄胎蛋白抗血清可阻止抗Arc-1与MDCK细胞的结合。Arc-1抗原的分布也与葡萄胎蛋白非常相似：它在MDCK细胞和各种犬组织中的细胞-细胞接触处都有丰富的分布。在肠上皮中，抗原可以进一步定位在连接复合体的区域。为了研究解离抗体的作用机制，将生长在Boyden小室核孔滤器上的MDCK细胞暴露于上下两个隔室的抗Arc-1。结果表明，该抗体仅从基底细胞表面干扰细胞黏附，而不从顶端细胞表面干扰细胞黏附。秋水仙素对抗体作用有抑制作用，而细胞松弛素B对抗体作用无抑制作用。此外，细胞内cAMP水平升高可防止细胞解离。这些发现表明，抗Arc-1抗体作用于紧密连接下方的靶点(可能作用于连接复合体中的抗原)，并证实细胞骨架和代谢因子积极参与维持连接完整性。
It has previously been shown that the monoclonal antibody anti-Arc-1 dissociates Madin-Darby canine kidney (MDCK) epithelial cells and changes their morphology in vitro (Imhof, B.A., H.P. Vollmers, S.L. Goodman, and W. Birchmeier, 1983, Cell, 35:667-675). In this article we demonstrate that the anti-Arc-1 antibody recognizes an uvomorulin-like molecule on MDCK cells, i.e., it immunoprecipitates an 84-kD protein fragment from a tryptic digest of cell surfaces in the presence of Ca2+ (as does anti-uvomorulin antiserum). Furthermore, anti-uvomorulin antiserum prevents the binding of anti-Arc-1 to MDCK cells. The distribution of the Arc-1 antigen is also quite similar to that of uvomorulin: it is enriched at the cell-cell contacts both of MDCK cells and of cells in various canine tissues. In the intestinal epithelium the antigen could be further localized in the region of the junctional complex. To study the mechanism of action of the dissociating antibody, MDCK cells grown on Nuclepore filters in Boyden chambers were exposed to anti-Arc-1 from either the upper or lower compartment. It could be shown that the antibody interfered with cell adhesion only from the basolateral but not from the apical cell surface. Antibody action was inhibited in the presence of colchicine but not cytochalasin B. Furthermore, cell dissociation was prevented when the cellular cAMP level was raised. These findings indicate that the anti-Arc-1 antibody acts on a target below the tight junctions (possibly on the antigen located in the junctional complex), and they confirm that cytoskeleton and metabolic factors are actively involved in the maintenance of junctional integrity.
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影响因子：
64.5
作者：
DAMSKY, CH;RICHA, J;BUCK, CA
通讯作者：
BUCK, CA
影响因子：
64.8
作者：
SADOUL, R;HIRN, M;GORIDIS, C
通讯作者：
GORIDIS, C
影响因子：
64.5
作者：
KREIS, TE;BIRCHMEIER, W
通讯作者：
BIRCHMEIER, W
DOI：
10.1083/jcb.100.1.327
发表时间：
1985-01
期刊：
The Journal of cell biology
影响因子：
--
作者：
Boller K;Vestweber D;Kemler R
通讯作者：
Kemler R
DOI：
10.1073/pnas.80.14.4384
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
EDELMAN, GM;GALLIN, WJ;THIERY, JP
通讯作者：
THIERY, JP