title: Procollagen C-proteinase enhancer grasps the stalk of the C-propeptide trimer to boost collagen precursor maturation.
abstract: Tight regulation of collagen fibril deposition in the extracellular matrix is essential for normal tissue homeostasis and repair, defects in which are associated with several degenerative or fibrotic disorders. A key regulatory step in collagen fibril assembly is the C-terminal proteolytic processing of soluble procollagen precursors. This step, carried out mainly by bone morphogenetic protein-1/tolloid-like proteinases, is itself subject to regulation by procollagen C-proteinase enhancer proteins (PCPEs) which can dramatically increase bone morphogenetic protein-1/tolloid-like proteinase activity, in a substrate-specific manner. Although it is known that this enhancing activity requires binding of PCPE to the procollagen C-propeptide trimer, identification of the precise binding site has so far remained elusive. Here, use of small-angle X-ray scattering provides structural data on this protein complex indicating that PCPE binds to the stalk region of the procollagen C-propeptide trimer, where the three polypeptide chains associate together, at the junction with the base region. This is supported by site-directed mutagenesis, which identifies two highly conserved, surface-exposed lysine residues in this region of the trimer that are essential for binding, thus revealing structural parallels with the interactions of Complement C1r/C1s, Uegf, BMP-1 (CUB) domain-containing proteins in diverse biological systems such as complement activation, receptor signaling, and transport. Together with detailed kinetics and interaction analysis, these results provide insights into the mechanism of action of PCPEs and suggest clear strategies for the development of novel antifibrotic therapies.
authors.affiliation: Unité Mixte de Recherche 5305, Institut de Biologie et Chimie des Protéines, Centre National de la Recherche Scientifique, Université Lyon 1, 69367 Lyon cedex 7, France.
authors.forename: Jean-Marie
authors.initials: JM
authors.lastname: Bourhis
authors.forename: Sandrine
authors.initials: S
authors.lastname: Vadon-Le Goff
authors.forename: Hassnae
authors.initials: H
authors.lastname: Afrache
authors.forename: Natacha
authors.initials: N
authors.lastname: Mariano
authors.forename: Daniel
authors.initials: D
authors.lastname: Kronenberg
authors.forename: Nicole
authors.initials: N
authors.lastname: Thielens
authors.forename: Catherine
authors.initials: C
authors.lastname: Moali
authors.forename: David J S
authors.initials: DJ
authors.lastname: Hulmes
mesh_terms: D000818:Animals; D001665:Binding Sites / Q000235:genetics; D055395:Bone Morphogenetic Protein 1 / Q000235:genetics / Q000378:metabolism*; D002850:Chromatography, Gel; D002942:Circular Dichroism; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism*; D053838:Scattering, Small Angle; D020349:Surface Plasmon Resonance
chemical_list: D024061:Collagen Type III; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; C491555:PCOLCE protein, human; D011994:Recombinant Proteins; C525705:BMP1 protein, human; D055395:Bone Morphogenetic Protein 1
references.citation: Nature. 2010 Mar 18;464(7287):445-8
references.pmid: 20237569
references.citation: Mol Cell Biol. 2006 Jan;26(1):238-49
references.pmid: 16354695
references.citation: Science. 1996 Jan 19;271(5247):360-2
references.pmid: 8553073
references.citation: J Biol Chem. 2007 Jun 8;282(23):16924-33
references.pmid: 17446170
references.citation: J Biol Chem. 2002 Dec 20;277(51):49820-30
references.pmid: 12393877
references.citation: Biophys J. 1999 Jun;76(6):2879-86
references.pmid: 10354416
references.citation: J Biol Chem. 2005 Jun 17;280(24):22616-23
references.pmid: 15817489
references.citation: Eur Biophys J. 2006 Sep;35(7):567-76
references.pmid: 16636827
references.citation: Int J Biochem Cell Biol. 2006 Mar;38(3):358-65
references.pmid: 16300990
references.citation: J Biol Chem. 2004 Oct 8;279(41):43336-44
references.pmid: 15292230
references.citation: J Immunol. 2009 Jan 1;182(1):456-65
references.pmid: 19109177
references.citation: Nat Struct Mol Biol. 2012 Oct;19(10):1031-6
references.pmid: 23001006
references.citation: J Biol Chem. 2008 Sep 12;283(37):25715-25724
references.pmid: 18596036
references.citation: Eur J Dermatol. 2009 Nov-Dec;19(6):552-64
references.pmid: 19625234
references.citation: J Biol Chem. 2011 Dec 9;286(49):41905-41911
references.pmid: 22027825
references.citation: Proc Natl Acad Sci U S A. 1996 May 14;93(10):5127-30
references.pmid: 8643539
references.citation: FEBS Lett. 2010 Feb 19;584(4):657-61
references.pmid: 20043912
references.citation: Am J Physiol Heart Circ Physiol. 2012 Jul 15;303(2):H234-40
references.pmid: 22610170
references.citation: Proteins. 2008 Mar;70(4):1154-61
references.pmid: 17932911
references.citation: J Biol Chem. 2001 Dec 28;276(52):48930-6
references.pmid: 11684689
references.citation: Biochem J. 2011 Oct 15;439(2):185-93
references.pmid: 21954942
references.citation: Eur J Biochem. 2001 May;268(10):2991-6
references.pmid: 11358517
references.citation: Proc Natl Acad Sci U S A. 2009 May 26;106(21):8561-6
references.pmid: 19429706
references.citation: EMBO J. 2004 Aug 4;23(15):3020-30
references.pmid: 15257288
references.citation: J Immunol. 2007 May 1;178(9):5710-6
references.pmid: 17442954
references.citation: J Biol Chem. 2004 Jul 9;279(28):29391-7
references.pmid: 15117939
references.citation: J Cell Sci. 2005 Apr 1;118(Pt 7):1341-53
references.pmid: 15788652
references.citation: Biophys J. 2001 Jun;80(6):2946-53
references.pmid: 11371467
references.citation: Curr Opin Cell Biol. 2008 Oct;20(5):495-501
references.pmid: 18640274
references.citation: J Biol Chem. 2003 Aug 22;278(34):32157-64
references.pmid: 12788922
references.citation: J Biol Chem. 2009 Jul 17;284(29):19340-8
references.pmid: 19473974
references.citation: Proc Natl Acad Sci U S A. 2012 Jul 31;109(31):12461-6
references.pmid: 22761315
references.citation: Coll Relat Res. 1986 Jul;6(3):267-77
references.pmid: 3533406
references.citation: J Biol Chem. 2009 Nov 27;284(48):33437-46
references.pmid: 19801683
references.citation: Exp Biol Med (Maywood). 2008 Feb;233(2):109-22
references.pmid: 18222966
references.citation: J Immunol. 2007 Jul 1;179(1):455-62
references.pmid: 17579066
references.citation: J Biol Chem. 2011 Nov 11;286(45):38932-8
references.pmid: 21940633
references.citation: J Appl Crystallogr. 2009 Apr 1;42(Pt 2):342-346
references.pmid: 27630371
references.citation: J Biol Chem. 2005 Jun 24;280(25):24188-94
references.pmid: 15834133
references.citation: Structure. 2011 Nov 9;19(11):1635-43
references.pmid: 22078562
references.citation: J Biol Chem. 2004 Apr 2;279(14):14065-73
references.pmid: 14724269
references.citation: Nat Med. 2012 Jul 06;18(7):1028-40
references.pmid: 22772564
references.citation: Hepatology. 1997 Sep;26(3):611-7
references.pmid: 9303490