Structure of recombinant N-terminal globule of type VI collagen α3 chain and its binding to heparin and hyaluronan

U. Specks, U. Mayer, R. Nischt, T. Spissinger, K. Mann, R. Timpl, J. Engel, M. L. Chu

Research output: Contribution to journalArticle

115 Scopus citations

Abstract

A large portion of the N-terminal globule of human collagen VI was prepared from the culture medium of stably transfected human embryonic kidney cell clones. The recombinant product corresponds to sequence positions 1-1586 of the α3(VI) chain that consists of eight homologous ~ 200 residue motifs (N9 to N2) being similar to the A domain motif of von Willebrand factor. By ultracentrifugation fragment N9-N2 showed a molecular mass of 180 kDa and an asymmetric shape. Elongated structures that consist of eight small globes (diameter ~ 5 nm) were demonstrated by electron microscopy. The data indicate that each A domain motif represents a separate folding unit which are connected to each other by short protease-sensitive peptide segments. Circular dichroism studies demonstrated about 38% α helix, 14% β sheets and 17% β turns. Fragment N9-N2 showed binding to heparin which could be abolished by moderate salt concentrations. Heparin binding was assigned to domains N9, N6 and N3 which were obtained after partial proteolysis. Domains N7, N5 and N4 lacked affinity for heparin. In addition, N9-N2 showed strong binding to hyaluronan that required exposure to 6 M urea for full dissociation. Ligand binding studies indicated some affinity of N9-N2 for the triple helical region of collagen VI suggesting a role of the N-terminal globule in the self-assembly of microfibrils. No or only little binding was, however, observed to fibril-forming collagens I and III, several basement membrane proteins and other extracellular proteins. Fragment N9-N2 was also an inactive substrate for cell adhesion.

Original languageEnglish (US)
Pages (from-to)4281-4290
Number of pages10
JournalEMBO Journal
Volume11
Issue number12
DOIs
StatePublished - 1992

Keywords

  • A domain motif
  • Collagen microfibrils
  • Eukaryotic expression vector
  • Glycosaminoglycans
  • Protein-protein and cell-matrix interactions

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

Fingerprint Dive into the research topics of 'Structure of recombinant N-terminal globule of type VI collagen α3 chain and its binding to heparin and hyaluronan'. Together they form a unique fingerprint.

  • Cite this