title: Properties of the collagenous domain of the alpha 3(IV) chain, the Goodpasture antigen, of lens basement membrane collagen. Selective cleavage of alpha (IV) chains with retention of their triple helical structure and noncollagenous domain.
abstract: A third chain, alpha 3(IV), of basement membrane collagen was recently discovered and was identified as the primary target for the autoantibodies of patients with Goodpasture syndrome (Saus, J., Wieslander, J., Langeveld, J. P. M., Quinones, S., and Hudson, B. G. (1988) J. Biol. Chem. 263, 13374-13380). In the present study, this chain was excised in the form of a truncated promoter by cleavage of basement membrane with Pseudomonas aeruginosa elastase and characterized. The triple helical structure and NC1 domain were retained. Elastase selectively cleaved at a site within the triple helical domain of the alpha 3 chain that is distinct from the cleavage site of the alpha 1 and alpha 2 chains. The truncated alpha 3 chain was found to contain 1460 residues, of which 1225 comprise the collagenous domain, and is cross-linked within this domain by disulfide bonds, forming a high Mr complex (greater than 300,000). Truncated protomers with a length of 340 nm corresponding to the theoretical length for the truncated alpha 3 chain were observed by electron microscopy as suprastructures in which the triple helical domains of three protomers were interwined. These protomers were also connected to each other and to the 140-nm protomers that appear to be comprised of the alpha 1 and alpha 2 chains. These results extended the known length of the alpha 3 chain by about 1000 residues and suggested that protomers of this chain self-associate through interactions between their triple helical domains and between their NC1 domains.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Kansas Medical Center, Kansas City 66103.
authors.forename: S
authors.initials: S
authors.lastname: Gunwar
authors.forename: M E
authors.initials: ME
authors.lastname: Noelken
authors.forename: B G
authors.initials: BG
authors.lastname: Hudson
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001324:Autoantigens / Q000737:chemistry*; D001485:Basement Membrane / Q000737:chemistry; D015153:Blotting, Western; D003094:Collagen / Q000737:chemistry* / Q000276:immunology; D024141:Collagen Type IV; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007908:Lens, Crystalline / Q000737:chemistry*; D051379:Mice; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D010196:Pancreatic Elastase / Q000494:pharmacology; D010446:Peptide Fragments / Q000737:chemistry; D011487:Protein Conformation
chemical_list: D000596:Amino Acids; D001324:Autoantigens; D024141:Collagen Type IV; D010446:Peptide Fragments; C057112:type IV collagen alpha3 chain; D003094:Collagen; D010196:Pancreatic Elastase