title: Mechanism of chain selection in the assembly of collagen IV: a prominent role for the alpha2 chain.
abstract: Collagens comprise a large superfamily of extracellular matrix proteins that play diverse roles in tissue function. The mechanism by which newly synthesized collagen chains recognize each other and assemble into specific triple-helical molecules is a fundamental question that remains unanswered. Emerging evidence suggests a role for the non-collagenous domain (NC1) located at the C-terminal end of each chain. In this study, we have investigated the molecular mechanism underlying chain selection in the assembly of collagen IV. Using surface plasmon resonance, we have determined the kinetics of interaction and assembly of the alpha1(IV) and alpha2(IV) NC1 domains. We show that the differential affinity of alpha2(IV) NC1 domain for dimer formation underlies the driving force in the mechanism of chain discrimination. Given its characteristic domain recognition and affinity for the alpha1(IV) NC1 domain, we conclude that the alpha2(IV) chain plays a regulatory role in directing chain composition in the assembly of (alpha1)(2)alpha2 triple-helical molecule. Detailed crystal structure analysis of the [(alpha1)(2)alpha2](2) NC1 hexamer and sequence alignments of the NC1 domains of all six alpha-chains from mammalian species revealed the residues involved in the molecular recognition of NC1 domains. We further identified a hypervariable region of 15 residues and a beta-hairpin structural motif of 13 residues as two prominent regions that mediate chain selection in the assembly of collagen IV. To our knowledge, this report is the first to combine kinetics and structural data to describe molecular basis for chain selection in the assembly of a collagen molecule.
authors.affiliation: Department of Medicine, Vanderbilt University School of Medicine, Medical Center, 1161 21st Avenue South, Nashville, TN 37232, USA. Jamshid.Khoshnoodi@vanderbilt.edu
authors.forename: Jamshid
authors.initials: J
authors.lastname: Khoshnoodi
authors.forename: Kristmundur
authors.initials: K
authors.lastname: Sigmundsson
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Cartailler
authors.forename: Olga
authors.initials: O
authors.lastname: Bondar
authors.forename: Munirathinam
authors.initials: M
authors.lastname: Sundaramoorthy
authors.forename: Billy G
authors.initials: BG
authors.lastname: Hudson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D024141:Collagen Type IV / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020836:Protein Structure, Quaternary; D016415:Sequence Alignment; D020349:Surface Plasmon Resonance
chemical_list: D024141:Collagen Type IV