A bacterial collagen-binding domain with novel calcium-binding motif controls domain orientation

Jeffrey J. Wilson, Osamu Matsushita, Akinobu Okabe, Joshua Sakon

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

The crystal structure of a collagen-binding domain (CBD) with an N-terminal domain linker from Clostridium histolyticum class I collagenase was determined at 1.00 Å resolution in the absence of calcium (1NQJ) and at 1.65 Å resolution in the presence of calcium (1NQD). The mature enzyme is composed of four domains: a metalloprotease domain, a spacing domain and two CBDs. A 12-residue-long linker is found at the N-terminus of each CBD. In the absence of calcium, the CBD reveals a β-sheet sandwich fold with the linker adopting an α-helix. The addition of calcium unwinds the linker and anchors it to the distal side of the sandwich as a new β-strand. The conformational change of the linker upon calcium binding is confirmed by changes in the Stokes and hydrodynamic radii as measured by size exclusion chromatography and by dynamic light scattering with and without calcium. Furthermore, extensive mutagenesis of conserved surface residues and collagen-binding studies allow us to identify the collagen-binding surface of the protein and propose likely collagen-protein binding models.

Original languageEnglish
Pages (from-to)1743-1752
Number of pages10
JournalEMBO Journal
Volume22
Issue number8
DOIs
Publication statusPublished - Apr 15 2003
Externally publishedYes

Fingerprint

Crystal orientation
Collagen
Calcium
Clostridium histolyticum
Clostridium
Mutagenesis
Size exclusion chromatography
Metalloproteases
Collagenases
Dynamic light scattering
Hydrodynamics
Anchors
Protein Binding
Gel Chromatography
Carrier Proteins
Membrane Proteins
Crystal structure
Enzymes

Keywords

  • Calcium-induced conformational change
  • Domain reorientation
  • Extracellular calcium-binding protein
  • Molecular switch
  • Square antiprismatic

ASJC Scopus subject areas

  • Genetics
  • Cell Biology

Cite this

A bacterial collagen-binding domain with novel calcium-binding motif controls domain orientation. / Wilson, Jeffrey J.; Matsushita, Osamu; Okabe, Akinobu; Sakon, Joshua.

In: EMBO Journal, Vol. 22, No. 8, 15.04.2003, p. 1743-1752.

Research output: Contribution to journalArticle

Wilson, Jeffrey J. ; Matsushita, Osamu ; Okabe, Akinobu ; Sakon, Joshua. / A bacterial collagen-binding domain with novel calcium-binding motif controls domain orientation. In: EMBO Journal. 2003 ; Vol. 22, No. 8. pp. 1743-1752.
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