COMPARATIVE-ANALYSIS OF THE NONCOLLAGENOUS NC1 DOMAIN OF TYPE-IV COLLAGEN - IDENTIFICATION OF STRUCTURAL FEATURES IMPORTANT FOR ASSEMBLY, FUNCTION, AND PATHOGENESIS

Type TV collagen alpha 1-alpha 6 chains have important roles in the as sembly of basement membranes and are implicated in the pathogenesis of Goodpasture syndrome, an autoimmune disorder, and Alport syndrome, a hereditary renal disease. We report comparative sequence analyses and structural predictions of the noncollagenous C-terminal globular NC 1 domain (28 sequences). The inferred tree verified that type IV collage n sequences fall into two groups, alpha 1-like and alpha 2-like, and s uggested that vertebrate alpha 3/alpha 4 sequences evolved before alph a 1/alpha 2 and alpha 5/alpha 6. About one fifth of NC1 residues were identified to confer either the alpha 1 or alpha 2 group-specificity. These residues accumulate opposite charge in subdomain B of alpha 1 (p ositive) and alpha 2 (negative) sequences and may play a role in the s toichiometric chain selection upon type TV collagen assembly. Neural n etwork secondary structure prediction on multiple aligned sequences re vealed a subdomain core structure consisting of six hydrophobic beta-s trands and one short alpha-helix with a significant hydrophobic moment . The existence of opposite charges in the alpha-helices may carry imp lications for intersubdomain interactions. The results provide a ratio nale for defining the epitope that binds Goodpasture autoantibodies an d a framework for understanding how certain NC1 mutations may lead to Alport syndrome. A search algorithm, based entirely on amino acid prop erties, yielded a possible similarity of NC1 to tissue inhibitor of me talloproteinases (TIMP) and prompted an investigation of a possible fu nctional relationship. The results indicate that NC1 preparations decr ease the activity of matrix metalloproteinases 2 and 3 (MMP-2, MMP-3) toward a peptide substrate, though not to [14C]-gelatin. We suggest th at an ancestral NC1 may have been incorporated into type TV collagen a s an evolutionarily mobile domain carrying proteinase inhibitor functi on.