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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

Authors

  • Weinacker A
  • Chen A
  • Agrez M
  • Cone RI
  • Nishimura S
  • Wayner E
  • Pytela R
  • Sheppard D

Journal

The Journal of biological chemistry

Publication Data

March 4, 1994

Associated Grants

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

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