title: A manganese-rich environment supports superoxide dismutase activity in a Lyme disease pathogen, Borrelia burgdorferi.
abstract: The Lyme disease pathogen Borrelia burgdorferi represents a novel organism in which to study metalloprotein biology in that this spirochete has uniquely evolved with no requirement for iron. Not only is iron low, but we show here that B. burgdorferi has the capacity to accumulate remarkably high levels of manganese. This high manganese is necessary to activate the SodA superoxide dismutase (SOD) essential for virulence. Using a metalloproteomic approach, we demonstrate that a bulk of B. burgdorferi SodA directly associates with manganese, and a smaller pool of inactive enzyme accumulates as apoprotein. Other metalloproteins may have similarly adapted to using manganese as co-factor, including the BB0366 aminopeptidase. Whereas B. burgdorferi SodA has evolved in a manganese-rich, iron-poor environment, the opposite is true for Mn-SODs of organisms such as Escherichia coli and bakers' yeast. These Mn-SODs still capture manganese in an iron-rich cell, and we tested whether the same is true for Borrelia SodA. When expressed in the iron-rich mitochondria of Saccharomyces cerevisiae, B. burgdorferi SodA was inactive. Activity was only possible when cells accumulated extremely high levels of manganese that exceeded cellular iron. Moreover, there was no evidence for iron inactivation of the SOD. B. burgdorferi SodA shows strong overall homology with other members of the Mn-SOD family, but computer-assisted modeling revealed some unusual features of the hydrogen bonding network near the enzyme's active site. The unique properties of B. burgdorferi SodA may represent adaptation to expression in the manganese-rich and iron-poor environment of the spirochete.
authors: Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland 21205.
authors: J Dafhne
authors: JD
authors: Aguirre
authors: Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland 21205.
authors: Hillary M
authors: HM
authors: Clark
authors: Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543.
authors: Matthew
authors: M
authors: McIlvin
authors: Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland 21205.
authors: Christine
authors: C
authors: Vazquez
authors: Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland 21205.
authors: Shaina L
authors: SL
authors: Palmere
authors: Department of Pathology, Division of Medical Microbiology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
authors: Dennis J
authors: DJ
authors: Grab
authors: Department of Biology, University of Texas, San Antonio, Texas 78249.
authors: J
authors: J
authors: Seshu
authors: Geriatric Research, Education, and Clinical Center, South Texas Veterans Health Care System, Department of Veterans Affairs, San Antonio, Texas 78229; Department of Biochemistry, University of Texas Health Science Center, San Antonio, Texas 78229.
authors: P John
authors: PJ
authors: Hart
authors: Marine Chemistry and Geochemistry, Woods Hole Oceanographic Institution, Woods Hole, Massachusetts 02543.
authors: Mak
authors: M
authors: Saito
authors: Department of Biochemistry and Molecular Biology, Johns Hopkins University Bloomberg School of Public Health, Baltimore, Maryland 21205. Electronic address: vculotta@jhsph.edu.
authors: Valeria C
authors: VC
authors: Culotta
mesh_terms: D000595:Amino Acid Sequence; D001051:Apoenzymes / Q000302:isolation & purification / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D025065:Borrelia burgdorferi / Q000201:enzymology*; D020134:Catalytic Domain; D017124:Conserved Sequence; D004789:Enzyme Activation; D006860:Hydrogen Bonding; D006861:Hydrogen Peroxide; D008345:Manganese / Q000378:metabolism / Q000502:physiology*; D008928:Mitochondria / Q000201:enzymology; D008958:Models, Molecular; D008969:Molecular Sequence Data; D021381:Protein Transport; D012441:Saccharomyces cerevisiae; D017386:Sequence Homology, Amino Acid; D013482:Superoxide Dismutase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D001051:Apoenzymes; D001426:Bacterial Proteins; D008345:Manganese; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase
references: Biochim Biophys Acta. 2010 Feb;1804(2):298-307
references: 19699328
references: J Mol Biol. 1993 Dec 5;234(3):779-815
references: 8254673
references: J Clin Chem Clin Biochem. 1985 Jul;23(7):393-8
references: 4045387
references: Acta Crystallogr Sect F Struct Biol Cryst Commun. 2011 Oct 1;67(Pt 10):1173-8
references: 22102021
references: Biochemistry. 1998 Apr 7;37(14):4722-30
references: 9537987
references: J Biol Chem. 2012 Apr 20;287(17):13524-31
references: 22389496
references: Mol Microbiol. 2002 Jun;44(5):1269-86
references: 12028379
references: Arch Biochem Biophys. 2012 Jul 15;523(2):191-7
references: 22561997
references: EMBO J. 2006 Apr 19;25(8):1775-83
references: 16601688
references: J Biol Chem. 1991 Jan 5;266(1):303-8
references: 1985901
references: Biochemistry. 2009 Apr 21;48(15):3417-24
references: 19265433
references: Mol Microbiol. 2009 Feb;71(3):594-612
references: 19040638
references: J Biol Chem. 2007 Feb 9;282(6):3837-46
references: 17148438
references: Infect Immun. 2011 Feb;79(2):732-44
references: 21078860
references: Infect Immun. 1997 Nov;65(11):4865-8
references: 9353077
references: Eur J Biochem. 1999 Aug;263(3):726-35
references: 10469136
references: J Biol Chem. 2001 Dec 14;276(50):47556-62
references: 11602606
references: Proc Natl Acad Sci U S A. 2012 Jan 3;109(1):95-100
references: 22198771
references: Cell Host Microbe. 2011 Aug 18;10(2):158-64
references: 21843872
references: Nature. 2010 Aug 5;466(7307):779-82
references: 20639861
references: Biochim Biophys Acta. 2010 Feb;1804(2):318-25
references: 19837190
references: J Biol Chem. 2009 Aug 21;284(34):22633-40
references: 19561359
references: Acc Chem Res. 2004 Jul;37(7):461-70
references: 15260508
references: Genetics. 2011 Dec;189(4):1261-70
references: 21926297
references: J Mol Biol. 2010 May 21;398(5):703-14
references: 20359484
references: J Biol Chem. 2012 Jun 1;287(23):19284-93
references: 22500025
references: Biochem Biophys Res Commun. 2012 Feb 10;418(2):414-9
references: 22281500
references: J Biol Inorg Chem. 2010 Sep;15(7):1051-62
references: 20429018
references: J Infect Dis. 2004 Jul 1;190(1):136-47
references: 15195253
references: Bioorg Chem. 2011 Apr;39(2):73-87
references: 21397291
references: Trends Microbiol. 2002 Nov;10(11):496-501
references: 12419613
references: Proc Natl Acad Sci U S A. 2003 Sep 2;100(18):10353-7
references: 12890866
references: Science. 2001 Jun 29;292(5526):2488-92
references: 11397910
references: Free Radic Biol Med. 2007 Aug 1;43(3):423-30
references: 17602958
references: Mol Microbiol. 2001 Jun;40(5):1175-86
references: 11401721
references: Curr Opin Chem Biol. 2010 Apr;14(2):218-24
references: 20015678
references: PLoS One. 2011;6(9):e24741
references: 21931840
references: Inflamm Res. 1998 Jul;47(7):277-84
references: 9719491
references: Proc Natl Acad Sci U S A. 2012 Sep 4;109(36):14314-9
references: 22908245
references: J Inorg Biochem. 2012 Jan;106(1):84-9
references: 22112844
references: PLoS One. 2010 Sep 03;5(9):e12570
references: 20838443
references: J Biol Chem. 1992 May 5;267(13):9140-5
references: 1577750
references: J Biol Chem. 2004 Jun 25;279(26):27339-44
references: 15082717
references: Genetics. 1989 May;122(1):19-27
references: 2659436
references: Science. 2000 Jun 2;288(5471):1651-3
references: 10834845
references: BMC Bioinformatics. 2011 Feb 28;12:64
references: 21356119
references: Methods Enzymol. 1984;105:93-104
references: 6328209
references: J Biol Chem. 2004 Mar 5;279(10):9248-54
references: 14672935
references: J Biol Chem. 2012 Apr 20;287(17):13541-8
references: 22247543
references: J Bacteriol. 1981 Jan;145(1):442-51
references: 6257639
references: Mol Microbiol. 2012 Dec;86(6):1441-51
references: 23061404
references: Genome Biol. 2005;6(9):R77
references: 16168084
references: Microbes Infect. 2012 Mar;14(3):207-16
references: 22033148
references: Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3449-54
references: 19218460
references: Curr Opin Biotechnol. 2012 Feb;23(1):89-95
references: 22138493
references: Semin Immunopathol. 2012 Mar;34(2):215-35
references: 22048835