Paolo Bonaldo, Vincenzo Russo, Francesco Bucciotti, Roberto Doliana, and Alfonso Colombatti* Divisione di Oncologia Sperimentale 2, Centro di Riferimento Oncologico, 33081 Aviano, Italy Received July 6, 1989; Revised Manuscript Received September 19, 1989 abstract: Type VI collagen is a component of 100 nm long periodic filaments with a widespread distribution around collagen fibers and on the surface of cells. It is an unusual collagen constituted by three distinct chains, one of which (a3) is much larger than the others and is encoded by a 9-kb mRNA. The amino acid sequence of the a3 (VI) deduced from the present cDNA clones specifies for a multidomain protein of at least 2648 residues made of a short collagenous sequence (336 residues), flanked at the N-terminus by nine 200 residue long repeating motifs and at the C-terminus by two similar motifs that share extensive identities with the collagen-binding type A repeats of von Willebrand factor. Type VI collagen and a3 (VI) fusion proteins bound to insolubilized type I collagen in a specific, time-dependent, and saturable manner. The a3 (VI) chain has three Arg-Gly-Asp sequences in the collagenous domain, and cell attachment was stimulated by the triple helix of type VI collagen and by a3 (VI) fusion proteins containing Arg-Gly-Asp sequences. This function was specifically inhibited by the Arg-Gly-Asp-Ser synthetic peptide. The type