Structural organization of distinct domains within the non-collagenous N-terminal region of collagen type XI

Kate E. Gregory, Julia T. Oxford, Yanwen Chen, Jay E. Gambee, Steven P. Gygi, Ruedi Aebersold, Peter J. Neame, Diane E. Mechling, Hans Peter Bächinger, Nicholas P. Morris

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

Collagen XI is a heterotrimeric molecule found predominantly in heterotypic cartilage fibrils, where it is involved in the regulation of fibrillogenesis. This function is thought to involve the complex N-terminal domain. The goal of this current study was to examine its structural organization to further elucidate the regulatory mechanism. The amino- propeptide (α1-Npp) alone or with isoforms of the variable region were recombinantly expressed and purified by affinity and molecular sieve chromatography. Cys-1-Cys-4 and Cys-2-Cys-3 disulfide bonds were detected by liquid chromatography-tandem mass spectrometry. This pattern is identical to the homologous α2-Npp, indicating that the recombinant proteins were folded correctly. Anomalous elution on molecular sieve chromatography suggested that the variable region was extended, which was confirmed using rotary shadowing; the α1-Npp formed a globular 'head' and the variable region an extended 'tail.' Circular dichroism spectra analysis determined that the α1-Npp comprised 33% β-sheet, whereas the variable region largely comprised non- periodic structure. Taken together, these results imply that the α1-Npp cannot be accommodated within the core of the fibril and that the variable region and]or minor helix facilitates its exclusion to the fibril surface. This provides further support for regulation of fibril diameter by steric hindrance or by interactions with other matrix components that affect fibrillogenesis.

Original languageEnglish (US)
Pages (from-to)11498-11506
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number15
DOIs
StatePublished - Apr 14 2000
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint

Dive into the research topics of 'Structural organization of distinct domains within the non-collagenous N-terminal region of collagen type XI'. Together they form a unique fingerprint.

Cite this