Structural comparison of ColH and ColG collagen-binding domains from Clostridium histolyticum

Ryan Bauer, Jeffrey J. Wilson, Sagaya Theresa Leena Philominathan, Dan Davis, Osamu Matsushita, Joshua Sakona

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Clostridium histolyticum secretes collagenases, ColG and ColH, that cause extensive tissue destruction in myonecrosis. The Cterminal collagen-binding domain (CBD) of collagenase is required for insoluble collagen fibril binding and subsequent collagenolysis. The high-resolution crystal structures of ColG-CBD (s3b) and ColH-CBD (s3) are reported in this paper. The new Xray structure of s3 was solved at 2.0-å resolution (R=17.4%; Rfree=23.3%), while the resolution of the previously determined s3b was extended to 1.4 å (R=17.9%; Rfree=21.0%). Despite sharing only 30% sequence identity, the molecules resemble one another closely (root mean square deviation [RMSD] Cα=1.5 å). All but one residue, whose side chain chelates with Ca2+, are conserved. The dual Ca2+ binding site in s3 is completed by an unconserved aspartate. Differential scanning calorimetric measurements showed that s3 gains thermal stability, comparable to s3b, by binding to Ca2+ (holo Tm=94.1°C; apo Tm=70.2°C). holo s3 is also stabilized against chemical denaturants urea and guanidine HCl. The three most critical residues for collagen interaction in s3b are conserved in s3. The general shape of the binding pocket is retained by altered loop structures and side chain positions. Small-angle X-ray scattering data revealed that s3 also binds asymmetrically to minicollagen. Besides the calciumbinding sites and the collagen-binding pocket, architecturally important hydrophobic residues and the hydrogen-bonding network around the cis-peptide bond are well conserved within the metallopeptidase subfamily M9B. CBDs were previously shown to bind to the extracellular matrix of various tissues. Compactness and extreme stability in physiological Ca2+ concentration possibly make both CBDs suitable for targeted growth factor delivery.

Original languageEnglish
Pages (from-to)318-327
Number of pages10
JournalJournal of Bacteriology
Volume195
Issue number2
DOIs
Publication statusPublished - Jan 2013

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Clostridium histolyticum
Collagen
Microbial Collagenase
Binding Sites
X-Rays
Guanidine
Metalloproteases
Collagenases
Hydrogen Bonding
Aspartic Acid
Extracellular Matrix
Urea
Intercellular Signaling Peptides and Proteins
Hot Temperature
Peptides

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Structural comparison of ColH and ColG collagen-binding domains from Clostridium histolyticum. / Bauer, Ryan; Wilson, Jeffrey J.; Philominathan, Sagaya Theresa Leena; Davis, Dan; Matsushita, Osamu; Sakona, Joshua.

In: Journal of Bacteriology, Vol. 195, No. 2, 01.2013, p. 318-327.

Research output: Contribution to journalArticle

Bauer, Ryan ; Wilson, Jeffrey J. ; Philominathan, Sagaya Theresa Leena ; Davis, Dan ; Matsushita, Osamu ; Sakona, Joshua. / Structural comparison of ColH and ColG collagen-binding domains from Clostridium histolyticum. In: Journal of Bacteriology. 2013 ; Vol. 195, No. 2. pp. 318-327.
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