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

J. Biol. Chem., Vol. 282, Issue 23, 16924-16933, June 8, 2007

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 282/23/16924    most recent M701610200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Blanc, G. Articles by Moali, C. Search for Related Content PubMed PubMed Citation Articles by Blanc, G. Articles by Moali, C. Social Bookmarking                      What's this?

Insights into How CUB Domains Can Exert Specific Functions while Sharing a Common Fold

CONSERVED AND SPECIFIC FEATURES OF THE CUB1 DOMAIN CONTRIBUTE TO THE MOLECULAR BASIS OF PROCOLLAGEN C-PROTEINASE ENHANCER-1 ACTIVITY^*

From the Institut de Biologie et Chimie des Protéines, CNRS/Université Claude Bernard Lyon 1, Unité Mixte de Recherche 5086, Institut Fédératif de Recherche 128 Biosciences Lyon Gerland, 7 Passage du Vercors, 69367 Lyon cedex 7, France

Procollagen C-proteinase enhancers (PCPE-1 and -2) are extracellular glycoproteins that can stimulate the C-terminal processing of fibrillar procollagens by tolloid proteinases such as bone morphogenetic protein-1. They consist of two CUB domains (CUB1 and -2) that alone account for PCPE-enhancing activity and one C-terminal NTR domain. CUB domains are found in several extracellular and plasma membrane-associated proteins, many of which are proteases. We have modeled the structure of the CUB1 domain of PCPE-1 based on known three-dimensional structures of CUB-containing proteins. Sequence alignment shows conserved amino acids, notably two acidic residues (Asp-68 and Asp-109) involved in a putative surface-located calcium binding site, as well as a conserved tyrosine residue (Tyr-67). In addition, three residues (Glu-26, Thr-89, and Phe-90) are found only in PCPE CUB1 domains, in putative surface-exposed loops. Among the conserved residues, it was found that mutations of Asp-68 and Asp-109 to alanine almost completely abolished PCPE-1 stimulating activity, whereas mutation of Tyr-67 led to a smaller reduction of activity. Among residues specific to PCPEs, mutation of Glu-26 and Thr-89 had little effect, whereas mutation of Phe-90 dramatically decreased the activity. Changes in activity were paralleled by changes in binding of different PCPE-1 mutants to a mini-procollagen III substrate, as shown by surface plasmon resonance. We conclude that PCPE-stimulating activity requires a calcium binding motif in the CUB1 domain that is highly conserved among CUB-containing proteins but also that PCPEs contain specific sites that could become targets for the development of novel anti-fibrotic therapies.

Received for publication, February 23, 2007 , and in revised form, April 12, 2007.

^* This work was supported by the Région Rhône-Alpes, the European Commission (Contract No. NMP2-CT-2003-504017), the Ligue Contre le Cancer, Engelhard (Lyon), the Centre National de la Recherche Scientifique and the Université Claude Bernard Lyon 1. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The on-line version of this article (available at http://www.jbc.org) contains supplemental Table S1 and Figs. S1 and S2.

¹ Present address: Commissariat à l'Energie Atomique-Grenoble/Département Réponse et Dynamique Cellulaires/Laboratoire de Biophysique Moléculaire et Cellulaire, CNRS Unité Mixte de Recherche 5090, 38054 Grenoble, France.

² To whom correspondence should be addressed. Tel.: 33-472-722-667; Fax: 33-472-722-604; E-mail: d.hulmes{at}ibcp.fr.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Zhu, J. Gardner, C. R. Pullinger, J. P. Kane, J. F. Thompson, and O. L. Francone Regulation of apoAI processing by procollagen C-proteinase enhancer-2 and bone morphogenetic protein-1 J. Lipid Res., July 1, 2009; 50(7): 1330 - 1339. [Abstract] [Full Text] [PDF]