The dissection of the activities mediated by type I collagen requires an approach by which the influence of triple-helical conformation can be evaluated. The and integrin binding sites within type I collagen are dependent upon triple-helical conformation and contained within residues 124-822 from 1(I). Seven 1(I)-derived triple-helical peptides (THPs) were synthesized based on charge clustering (1(I)256-270, 1(I)385-396, 1(I)406-417, 1(I)415-423, 1(I)448-456, 1(I)496-507, and 1(I)526-537). Three additional THPs were synthesized (1(I)85-96, 1(I)433-441, and 1(I)772-786) based on previously described or proposed activities (Kleinman, H. K., McGoodwin, E. B., Martin, G. R., Klebe, R. J., Fietzek, P. P., and Wooley, D. E.(1978) J. Biol. Chem. 253, 5642-5646; Staatz, W. D., Fok, K. F., Zutter, M. M., Adams, S. P., Rodriguez, B. A., and Santoro, S. A.(1991) J. Biol. Chem. 266, 7363-7367; San Antonio, J. D., Lander, A. D., Karnovsky, M. J., and Slayter, H. S.(1994) J. Cell Biol. 125, 1179-1188). Of the ten THPs, 1(I)772-786 THP had the greatest activity, with half-maximal normal dermal fibroblast adhesion occurring at a peptide concentration of 1.6 µM. Triple-helicity was essential for activity of this sequence, as the non-triple-helical peptide analog (1(I)772-786 SSP) exhibited considerably lower levels of cell adhesion promotion even at peptide concentrations as high as 100 µM. Within the sequence itself, residues 784-786 (Gly-Leu-Hyp) were important for cellular recognition, as the 1(I)772-783 THP had greatly reduced cell adhesion activity compared with 1(I)772-786 THP. Preliminary studies indicate that the integrin subunit mediates fibroblast adhesion to 1(I)772-786 THP. The identification of fibroblast integrin binding sites within type I collagen may have important implications for understanding collagen metabolism.

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