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Role of the integrin alpha v beta 6 in cell attachment to fibronectin. Heterologous expression of intact and secreted forms of the receptor.

The integrin alpha v beta 6 has been shown to be a fibronectin-binding protein. To determine whether the cytoplasmic and transmembrane domains of alpha v beta 6 are necessary for binding to fibronectin, a truncated, secreted form of the integrin lacking these domains was engineered and expressed in Chinese hamster ovary cells. Fibronectin affinity chromatography demonstrated that the secreted integrin, like its full-length counterpart, was capable of binding fibronectin. Monoclonal antibodies were made to secreted alpha v beta 6 and to beta 6-transfected NIH 3T3 cells. In experiments designed to determine whether alpha v beta 6 can mediate cell attachment to fibronectin, full-length human beta 6 was expressed in Chinese hamster ovary cells and in the human colon carcinoma cell line SW480. beta 6-expressing cells were identified by alpha v beta 6-specific antibodies, and the beta 6-transfectants were used in cell-adhesion assays. In Chinese hamster ovary cells, human beta 6 associated with hamster alpha v but was incapable of mediating cell attachment to fibronectin. However, expression of beta 6 in these cells had the dominant negative effect of decreasing alpha v beta 5-dependent adhesion to vitronectin. In SW480 cells, beta 6 expression conferred the ability to bind to fibronectin even in the presence of inhibitory antibodies against beta 1 integrins. In such cells, fibronectin binding ability could be blocked by an antibody to alpha v integrins. These results constitute the first direct evidence that alpha v beta 6 mediates cell attachment to fibronectin.

Pubmed ID: 8120056 RIS Download

Mesh terms: 3T3 Cells | Animals | Antibodies, Monoclonal | Antigens, Neoplasm | CHO Cells | Cell Adhesion | Cell Line | Chromatography, Affinity | Cricetinae | Fibronectins | Flow Cytometry | Gene Expression | HeLa Cells | Humans | Integrins | Mice | Mice, Inbred BALB C | Molecular Weight | Plasmids | Recombinant Proteins | Transfection

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Associated grants

  • Agency: NHLBI NIH HHS, Id: HL/A133259
  • Agency: NHLBI NIH HHS, Id: HL25816
  • Agency: NHLBI NIH HHS, Id: HL47412

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