Substrate Recognition by the Collagen-binding Domain of Clostridium histolyticum Class I Collagenase

Osamu Matsushita, Takaki Koide, Ryoji Kobayashi, Kazuhiro Nagata, Akinobu Okabe

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

Clostridium histolyticum type I collagenase (ColG) has a segmental structure, S1+S2+S3a+S3b. S3a and S3b bound to insoluble collagen, but S2 did not, thus indicating that S3 forms a collagen-binding domain (CBD). Because S3a+S3b showed the most efficient binding to substrate, cooperative binding by both domains was suggested for the enzyme. Monomeric (S3b) and tandem (S3a+S3b) CBDs bound to atelocollagen, which contains only the collagenous region. However, they did not bind to telopeptides immobilized on Sepharose beads. These results suggested that the binding site(s) for the CBD is(are) present in the collagenous region. The CBD bound to immobilized collagenous peptides, (Pro-Hyp-Gly)n and (Pro-Pro-Gly)n, only when n is large enough to allow the peptides to have a triple-helical conformation. They did not bind to various peptides with similar amino acid sequences or to gelatin, which lacks a triple-helical conformation. The CBD did not bind to immobilized Glc-Gal disaccharide, which is attached to the side chains of hydroxylysine residues in the collagenous region. These observations suggested that the CBD specifically recognizes the triple-helical conformation made by three polypeptide chains in the collagenous region.

Original languageEnglish
Pages (from-to)8761-8770
Number of pages10
JournalJournal of Biological Chemistry
Volume276
Issue number12
DOIs
Publication statusPublished - 2001 Mar 23
Externally publishedYes

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Clostridium histolyticum
Clostridium
Collagenases
Collagen
Substrates
Conformations
Peptides
prolyl-prolyl-glycine
Microbial Collagenase
Hydroxylysine
Disaccharides
Gelatin
Sepharose
Amino Acid Sequence
Binding Sites
Amino Acids
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Substrate Recognition by the Collagen-binding Domain of Clostridium histolyticum Class I Collagenase. / Matsushita, Osamu; Koide, Takaki; Kobayashi, Ryoji; Nagata, Kazuhiro; Okabe, Akinobu.

In: Journal of Biological Chemistry, Vol. 276, No. 12, 23.03.2001, p. 8761-8770.

Research output: Contribution to journalArticle

Matsushita, Osamu ; Koide, Takaki ; Kobayashi, Ryoji ; Nagata, Kazuhiro ; Okabe, Akinobu. / Substrate Recognition by the Collagen-binding Domain of Clostridium histolyticum Class I Collagenase. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 12. pp. 8761-8770.
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