title: Solution structure of clostridial collagenase H and its calcium-dependent global conformation change.
abstract: Collagenase H (ColH) from Clostridium histolyticum is a multimodular protein composed of a collagenase module (activator and peptidase domains), two polycystic kidney disease-like domains, and a collagen-binding domain. The interdomain conformation and its changes are very important for understanding the functions of ColH. In this study, small angle x-ray scattering and limited proteolysis were employed to reveal the interdomain arrangement of ColH in solution. The ab initio beads model indicated that ColH adopted a tapered shape with a swollen head. Under calcium-chelated conditions (with EGTA), the overall structure was further elongated. The rigid body model indicated that the closed form of the collagenase module was preferred in solution. The limited proteolysis demonstrated that the protease sensitivity of ColH was significantly increased under the calcium-chelated conditions, and that the digestion mainly occurred in the domain linker regions. Fluorescence measurements with a fluorescent dye were performed with the limited proteolysis products after separation. The results indicated that the limited proteolysis products exhibited fluorescence similar to that of the full-length ColH. These findings suggested that the conformation of full-length ColH in solution is the elongated form, and this form is calcium-dependently maintained at the domain linker regions.
authors.affiliation: Graduate School of Biomedical Engineering, Tohoku University, Sendai, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Ohbayashi
authors.forename: Takashi
authors.initials: T
authors.lastname: Matsumoto
authors.forename: Hiroki
authors.initials: H
authors.lastname: Shima
authors.forename: Masafumi
authors.initials: M
authors.lastname: Goto
authors.forename: Kimiko
authors.initials: K
authors.lastname: Watanabe
authors.forename: Akihito
authors.initials: A
authors.lastname: Yamano
authors.forename: Yasutake
authors.initials: Y
authors.lastname: Katoh
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Igarashi
authors.forename: Youhei
authors.initials: Y
authors.lastname: Yamagata
authors.forename: Kazutaka
authors.initials: K
authors.lastname: Murayama
mesh_terms: D002118:Calcium / Q000378:metabolism*; D047009:Hathewaya histolytica / Q000201:enzymology; D017364:Collagenases / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D011487:Protein Conformation; D059748:Proteolysis; D053838:Scattering, Small Angle; D012996:Solutions; D014961:X-Ray Diffraction
chemical_list: D012996:Solutions; D017364:Collagenases; D002118:Calcium
references.citation: J Am Soc Mass Spectrom. 2012 Mar;23(3):505-19
references.pmid: 22207568
references.citation: Proc Natl Acad Sci U S A. 2003 Feb 4;100(3):1316-21
references.pmid: 12552129
references.citation: Structure. 2006 Apr;14(4):653-60
references.pmid: 16615907
references.citation: J Biol Chem. 1998 Feb 6;273(6):3643-8
references.pmid: 9452493
references.citation: J Struct Biol. 2010 Oct;172(1):128-41
references.pmid: 20558299
references.citation: Biol Chem. 2010 Jul;391(7):737-43
references.pmid: 20482320
references.citation: J Biol Chem. 2001 Mar 23;276(12):8761-70
references.pmid: 11121400
references.citation: FEBS J. 2009 Jul;276(13):3589-601
references.pmid: 19490118
references.citation: Biol Chem. 2011 Nov;392(11):1039-45
references.pmid: 21871007
references.citation: BMC Struct Biol. 2012 Jul 16;12:17
references.pmid: 22800408
references.citation: Biophys J. 2005 Aug;89(2):1237-50
references.pmid: 15923225
references.citation: Appl Environ Microbiol. 2012 Aug;78(16):5839-44
references.pmid: 22685155
references.citation: Anal Biochem. 2001 Aug 1;295(1):101-6
references.pmid: 11476550
references.citation: Protein Sci. 2010 Apr;19(4):642-57
references.pmid: 20120026
references.citation: Bioinformatics. 2006 Jan 15;22(2):195-201
references.pmid: 16301204
references.citation: Nucleic Acids Res. 2010 Jul;38(Web Server issue):W563-8
references.pmid: 20507913
references.citation: Biophys J. 2001 Jun;80(6):2946-53
references.pmid: 11371467
references.citation: Biotechniques. 1992 Jun;12(6):870-4, 876-9
references.pmid: 1642895
references.citation: J Biol Chem. 2009 Apr 17;284(16):10868-76
references.pmid: 19208618
references.citation: J Bacteriol. 2013 Jan;195(2):318-27
references.pmid: 23144249
references.citation: J Appl Crystallogr. 2009 Apr 1;42(Pt 2):342-346
references.pmid: 27630371
references.citation: Biochemistry. 1984 Jun 19;23(13):3085-91
references.pmid: 6087888
references.citation: EMBO J. 2003 Apr 15;22(8):1743-52
references.pmid: 12682007
references.citation: Nucleic Acids Res. 1994 Nov 11;22(22):4673-80
references.pmid: 7984417
references.citation: Nat Struct Mol Biol. 2011 Sep 25;18(10):1109-14
references.pmid: 21947205