HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 276, Issue 12, 8761-8770, March 23, 2001

This Article Full Text Full Text (PDF) All Versions of this Article: 276/12/8761    most recent M003450200v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Matsushita, O. Articles by Okabe, A. Search for Related Content PubMed PubMed Citation Articles by Matsushita, O. Articles by Okabe, A. Social Bookmarking                      What's this?

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

Osamu Matsushita^§, Takaki Koide^¶**, Ryoji Kobayashi, Kazuhiro Nagata^¶, and Akinobu Okabe

From the Departments of  Microbiology and  Chemistry, Faculty of Medicine, Kagawa Medical University, Miki-cho, Kita-gun, Kagawa 761-0793, Japan, ^¶ Core Research for Evolutional Science and Technology, Japan Science and Technology Corporation, Kawaguchi, Saitama 332-0012, Japan, and the  Department of Molecular and Cellular Biology, Institute for Frontier Medical Sciences, Kyoto University, Kyoto 606-8397, Japan

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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EMBL Data Bank with accession number(s) .

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				J. C. Leo, H. Elovaara, B. Brodsky, M. Skurnik, and A. Goldman The Yersinia adhesin YadA binds to a collagenous triple-helical conformation but without sequence specificity Protein Eng. Des. Sel., August 1, 2008; 21(8): 475 - 484. [Abstract] [Full Text] [PDF]

				Y. Itoi, M. Horinaka, Y. Tsujimoto, H. Matsui, and K. Watanabe Characteristic Features in the Structure and Collagen-Binding Ability of a Thermophilic Collagenolytic Protease from the Thermophile Geobacillus collagenovorans MO-1. J. Bacteriol., September 1, 2006; 188(18): 6572 - 6579. [Abstract] [Full Text] [PDF]

				A. R. Tzafriri, M. Bercovier, and H. Parnas Reaction Diffusion Model of the Enzymatic Erosion of Insoluble Fibrillar Matrices Biophys. J., August 1, 2002; 83(2): 776 - 793. [Abstract] [Full Text] [PDF]

				Y. Hu, E. Webb, J. Singh, B. A. Morgan, J. A. Gainor, T. D. Gordon, and T. J. Siahaan Rapid Determination of Substrate Specificity of Clostridium histolyticumbeta -Collagenase Using an Immobilized Peptide Library J. Biol. Chem., March 1, 2002; 277(10): 8366 - 8371. [Abstract] [Full Text] [PDF]