title: The substrate-free and -bound crystal structures of the duplicated taurocyamine kinase from the human parasite Schistosoma mansoni.
abstract: The taurocyamine kinase from the blood fluke Schistosoma mansoni (SmTK) belongs to the phosphagen kinase (PK) family and catalyzes the reversible Mg(2+)-dependent transfer of a phosphoryl group between ATP and taurocyamine. SmTK is derived from gene duplication, as are all known trematode TKs. Our crystallographic study of SmTK reveals the first atomic structure of both a TK and a PK with a bilobal structure. The two unliganded lobes present a canonical open conformation and interact via their respective C- and N-terminal domains at a helix-mediated interface. This spatial arrangement differs from that observed in true dimeric PKs, in which both N-terminal domains make contact. Our structures of SmTK complexed with taurocyamine or l-arginine compounds explain the mechanism by which an arginine residue of the phosphagen specificity loop is crucial for substrate specificity. An SmTK crystal was soaked with the dead end transition state analog (TSA) components taurocyamine-NO3 (2-)-MgADP. One SmTK monomer was observed with two bound TSAs and an asymmetric conformation, with the first lobe semiclosed and the second closed. However, isothermal titration calorimetry and enzyme kinetics experiments showed that the two lobes function independently. A small angle x-ray scattering model of SmTK-TSA in solution with two closed active sites was generated.
authors.affiliation: From the Institut de Biologie et Chimie des Protéines, BMSSI-IBCP, UMR 5086 CNRS Université Lyon 1, 7, Passage du Vercors, 69367 Lyon Cedex 07, France and.
authors.forename: Romain
authors.initials: R
authors.lastname: Merceron
authors.affiliation: the Institut de Chimie et Biochimie Moléculaire et Supramoléculaire, UMR 5246 CNRS Université Lyon 1, 69622 Villeurbanne, France.
authors.forename: Ayman M
authors.initials: AM
authors.lastname: Awama
authors.affiliation: From the Institut de Biologie et Chimie des Protéines, BMSSI-IBCP, UMR 5086 CNRS Université Lyon 1, 7, Passage du Vercors, 69367 Lyon Cedex 07, France and.
authors.forename: Roland
authors.initials: R
authors.lastname: Montserret
authors.affiliation: the Institut de Chimie et Biochimie Moléculaire et Supramoléculaire, UMR 5246 CNRS Université Lyon 1, 69622 Villeurbanne, France olivier.marcillat@univ-lyon1.fr.
authors.forename: Olivier
authors.initials: O
authors.lastname: Marcillat
authors.affiliation: From the Institut de Biologie et Chimie des Protéines, BMSSI-IBCP, UMR 5086 CNRS Université Lyon 1, 7, Passage du Vercors, 69367 Lyon Cedex 07, France and p.gouet@ibcp.fr.
authors.forename: Patrice
authors.initials: P
authors.lastname: Gouet
mesh_terms: D000818:Animals; D018360:Crystallography, X-Ray; D015801:Helminth Proteins / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D017852:Phosphotransferases (Nitrogenous Group Acceptor) / Q000737:chemistry*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012550:Schistosoma mansoni / Q000201:enzymology*; D013654:Taurine / Q000031:analogs & derivatives* / Q000737:chemistry
chemical_list: D015801:Helminth Proteins; D013654:Taurine; C009558:taurocyamine; D017852:Phosphotransferases (Nitrogenous Group Acceptor); C022716:taurocyamine kinase
keywords: Schistosoma mansoni; isothermal titration calorimetry (ITC); small angle x-ray scattering (SAXS); substrate specificity; taurocyamine kinase; transition state; x-ray crystallography
references.citation: FEBS Lett. 1999 Nov 12;461(1-2):111-4
references.pmid: 10561506
references.citation: Protein Sci. 1999 Nov;8(11):2258-69
references.pmid: 10595529
references.citation: Proteins. 2000 May 15;39(3):216-25
references.pmid: 10737943
references.citation: Physiol Rev. 2000 Jul;80(3):1107-213
references.pmid: 10893433
references.citation: Acta Crystallogr D Biol Crystallogr. 2001 Feb;57(Pt 2):187-93
references.pmid: 11173463
references.citation: Annu Rev Physiol. 2001;63:289-325
references.pmid: 11181958
references.citation: Acta Crystallogr D Biol Crystallogr. 2001 Aug;57(Pt 8):1196-200
references.pmid: 11517911
references.citation: Biochemistry. 2002 Jun 4;41(22):6995-7000
references.pmid: 12033932
references.citation: Int J Biochem Cell Biol. 2002 Oct;34(10):1221-9
references.pmid: 12127572
references.citation: Biochemistry. 2002 Nov 26;41(47):13861-7
references.pmid: 12437342
references.citation: J Mol Graph Model. 2002 Dec;21(3):181-3
references.pmid: 12463636
references.citation: Protein Sci. 2003 Jan;12(1):103-11
references.pmid: 12493833
references.citation: J Exp Biol. 2003 May;206(Pt 9):1545-56
references.pmid: 12654893
references.citation: J Biol Chem. 2003 Jul 18;278(29):26952-7
references.pmid: 12732621
references.citation: Biochem J. 1958 Mar;68(3):447-52
references.pmid: 13522643
references.citation: Biochemistry. 2003 Nov 25;42(46):13596-604
references.pmid: 14622006
references.citation: Protein Sci. 2004 Mar;13(3):575-85
references.pmid: 14978299
references.citation: Nucleic Acids Res. 2004 Jul 1;32(Web Server issue):W668-73
references.pmid: 15215473
references.citation: Arch Biochem Biophys. 2007 Feb 15;458(2):158-66
references.pmid: 17239811
references.citation: Proteins. 2007 Oct 1;69(1):209-12
references.pmid: 17623863
references.citation: J Mol Biol. 2007 Sep 21;372(3):774-97
references.pmid: 17681537
references.citation: FEBS J. 2008 Feb;275(4):646-54
references.pmid: 18190534
references.citation: Methods Mol Biol. 2008;426:419-35
references.pmid: 18542881
references.citation: Comp Biochem Physiol B Biochem Mol Biol. 2008 Oct;151(2):176-82
references.pmid: 18639645
references.citation: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2008 Sep 1;64(Pt 9):854-7
references.pmid: 18765922
references.citation: FEBS Lett. 2008 Nov 26;582(28):3959-65
references.pmid: 18977227
references.citation: FEBS Lett. 2009 Jul 7;583(13):2218-24
references.pmid: 19500582
references.citation: Acta Crystallogr D Biol Crystallogr. 2009 Oct;65(Pt 10):1074-80
references.pmid: 19770504
references.citation: FASEB J. 2010 Jan;24(1):242-52
references.pmid: 19783784
references.citation: Structure. 2009 Oct 14;17(10):1356-67
references.pmid: 19836335
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
references.pmid: 20057044
references.citation: Biochemistry. 2010 Mar 9;49(9):2031-41
references.pmid: 20121101
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):133-44
references.pmid: 20124693
references.citation: Acta Crystallogr D Biol Crystallogr. 2010 Apr;66(Pt 4):486-501
references.pmid: 20383002
references.citation: Nucleic Acids Res. 2010 Jul;38(Web Server issue):W545-9
references.pmid: 20457744
references.citation: J Mol Biol. 2011 Jan 14;405(2):479-96
references.pmid: 21075117
references.citation: J Biol Chem. 2011 Mar 18;286(11):9338-50
references.pmid: 21212263
references.citation: Korean J Parasitol. 2010 Dec;48(4):281-4
references.pmid: 21234228
references.citation: Methods. 2011 Sep;55(1):94-106
references.pmid: 21821126
references.citation: Biochim Biophys Acta. 2012 Mar;1820(3):212-7
references.pmid: 21985891
references.citation: FEBS J. 2012 Aug;279(16):2863-75
references.pmid: 22715856
references.citation: Biochem Biophys Res Commun. 2012 Oct 12;427(1):212-7
references.pmid: 22995310
references.citation: J Biol Chem. 1990 Apr 25;265(12):6582-8
references.pmid: 2324092
references.citation: Int J Biol Macromol. 2013 Jun;57:273-7
references.pmid: 23523720
references.citation: Mol Biochem Parasitol. 2013 Apr;188(2):91-8
references.pmid: 23603791
references.citation: J Bioenerg Biomembr. 2013 Dec;45(6):511-8
references.pmid: 23873077
references.citation: PLoS Negl Trop Dis. 2013 Nov 21;7(11):e2548
references.pmid: 24278491
references.citation: Nucleic Acids Res. 2014 Jul;42(Web Server issue):W320-4
references.pmid: 24753421
references.citation: Mol Biochem Parasitol. 2014 Mar-Apr;194(1-2):56-63
references.pmid: 24815317
references.citation: Biochemistry. 2015 Jan 13;54(1):83-95
references.pmid: 25347386
references.citation: J Appl Crystallogr. 2009 Apr 1;42(Pt 2):342-346
references.pmid: 27630371
references.citation: Arch Biochem Biophys. 1986 Dec;251(2):558-66
references.pmid: 3800385
references.citation: Biochim Biophys Acta. 1987 Feb 11;890(2):233-41
references.pmid: 3801463
references.citation: Biopolymers. 1983 Dec;22(12):2577-637
references.pmid: 6667333
references.citation: J Biol Chem. 1980 Sep 10;255(17):8221-8
references.pmid: 7190972
references.citation: Mol Biochem Parasitol. 1994 Dec;68(2):319-22
references.pmid: 7739678
references.citation: Nature. 1996 May 23;381(6580):341-5
references.pmid: 8692275
references.citation: Biochim Biophys Acta. 1997 Dec 5;1343(2):152-9
references.pmid: 9434106
references.citation: Biochim Biophys Acta. 1976 Sep 14;445(2):364-76
references.pmid: 953037
references.citation: Proc Natl Acad Sci U S A. 1998 Jul 21;95(15):8449-54
references.pmid: 9671698
references.citation: Biophys J. 1998 Aug;75(2):1016-23
references.pmid: 9675202
references.citation: Biochim Biophys Acta. 1998 Oct 14;1388(1):253-9
references.pmid: 9774741
references.citation: FEBS Lett. 1998 Nov 13;439(1-2):133-7
references.pmid: 9849893