title: Ohr (organic hydroperoxide resistance protein) possesses a previously undescribed activity, lipoyl-dependent peroxidase.
abstract: The Ohr (organic hydroperoxide resistance) family of 15-kDa Cys-based, thiol-dependent peroxidases is central to the bacterial response to stress induced by organic hydroperoxides but not by hydrogen peroxide. Ohr has a unique three-dimensional structure and requires dithiols, but not monothiols, to support its activity. However, the physiological reducing system of Ohr has not yet been identified. Here we show that lipoylated enzymes present in the bacterial extracts of Xylella fastidiosa interacted physically and functionally with this Cys-based peroxidase, whereas thioredoxin and glutathione systems failed to support Ohr peroxidase activity. Furthermore, we could reconstitute in vitro three lipoyl-dependent systems as the Ohr physiological reducing systems. We also showed that OsmC from Escherichia coli, an orthologue of Ohr from Xylella fastidiosa, is specifically reduced by lipoyl-dependent systems. These results represent the first description of a Cys-based peroxidase that is directly reduced by lipoylated enzymes.
authors.affiliation: Departamento de Genética e Biologia Evolutiva, Instituto de Biociencias, Universidade de Sao Paulo, Rua do Matão 277, São Paulo 05508-900, Brazil.
authors.forename: José R R
authors.initials: JR
authors.lastname: Cussiol
authors.forename: Thiago G P
authors.initials: TG
authors.lastname: Alegria
authors.forename: Luke I
authors.initials: LI
authors.lastname: Szweda
authors.forename: Luis E S
authors.initials: LE
authors.lastname: Netto
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000378:metabolism; D007700:Kinetics; D054878:Lipoylation; D008954:Models, Biological; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000378:metabolism*; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D013379:Substrate Specificity; D044167:Xylella / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; C099394:osmC protein, E coli; C122138:peroxide repressor proteins; D010544:Peroxidases
references.citation: Antioxid Redox Signal. 2008 Sep;10(9):1515-26
references.pmid: 18500926
references.citation: Free Radic Biol Med. 2005 Jun 15;38(12):1543-52
references.pmid: 15917183
references.citation: J Biol Chem. 2004 Jun 11;279(24):25830-7
references.pmid: 15054099
references.citation: Proc Natl Acad Sci U S A. 2009 Apr 14;106(15):6226-31
references.pmid: 19339495
references.citation: Annu Rev Phytopathol. 1996;34:131-51
references.pmid: 15012538
references.citation: Toxicol Appl Pharmacol. 2002 Jul 1;182(1):84-90
references.pmid: 12127266
references.citation: Microbiology (Reading). 2001 Jul;147(Pt 7):1775-1782
references.pmid: 11429455
references.citation: Cell. 2007 Sep 7;130(5):797-810
references.pmid: 17803904
references.citation: J Biol Chem. 2003 Mar 28;278(13):11570-8
references.pmid: 12540833
references.citation: J Bacteriol. 1996 Jul;178(13):3695-700
references.pmid: 8682768
references.citation: Methods Mol Biol. 1994;32:9-15
references.pmid: 7951753
references.citation: J Biol Chem. 1997 Jul 18;272(29):17903-6
references.pmid: 9218413
references.citation: J Bacteriol. 1988 Sep;170(9):4315-21
references.pmid: 3137215
references.citation: Subcell Biochem. 2007;44:83-113
references.pmid: 18084891
references.citation: J Mol Biol. 2006 Jun 2;359(2):433-45
references.pmid: 16631787
references.citation: Microbes Infect. 2003 May;5(6):527-34
references.pmid: 12758282
references.citation: Plant Physiol. 2003 Aug;132(4):2045-57
references.pmid: 12913160
references.citation: Protein Sci. 2003 Dec;12(12):2838-43
references.pmid: 14627744
references.citation: J Biol Chem. 1958 May;232(1):143-58
references.pmid: 13549405
references.citation: Science. 2003 Oct 17;302(5644):462-4
references.pmid: 14564012
references.citation: Arch Biochem Biophys. 2005 Jan 1;433(1):240-54
references.pmid: 15581580
references.citation: EMBO J. 2002 Dec 16;21(24):6649-59
references.pmid: 12485986
references.citation: Acta Crystallogr D Biol Crystallogr. 2004 May;60(Pt 5):903-11
references.pmid: 15103136
references.citation: J Biol Chem. 2003 Dec 26;278(52):52834-40
references.pmid: 14551198
references.citation: Drug Metab Dispos. 2001 Jun;29(6):855-62
references.pmid: 11353754
references.citation: Cell. 1978 May;14(1):179-90
references.pmid: 352533
references.citation: J Bacteriol. 1983 Apr;154(1):344-50
references.pmid: 6339477
references.citation: Eur J Biochem. 2003 Mar;270(6):1036-42
references.pmid: 12631263
references.citation: Plant J. 2002 Oct;32(1):1-12
references.pmid: 12366796
references.citation: J Mol Biol. 1991 Aug 20;220(4):959-73
references.pmid: 1715407
references.citation: Proc Natl Acad Sci U S A. 2006 Jun 6;103(23):8662-7
references.pmid: 16735476
references.citation: Proc Natl Acad Sci U S A. 1995 May 9;92(10):4158-63
references.pmid: 11607542
references.citation: Methods Enzymol. 1994;234:454-61
references.pmid: 7808319
references.citation: J Bacteriol. 1995 Jan;177(1):1-10
references.pmid: 8002607
references.citation: Science. 2002 Feb 8;295(5557):1073-7
references.pmid: 11799204
references.citation: Antioxid Redox Signal. 2008 Sep;10(9):1565-76
references.pmid: 18498222
references.citation: J Biol Chem. 2001 Oct 19;276(42):38329-36
references.pmid: 11477096
references.citation: J Bacteriol. 1993 Mar;175(5):1325-36
references.pmid: 8444795
references.citation: Proteomics. 2003 Feb;3(2):224-37
references.pmid: 12601815
references.citation: J Biol Chem. 2009 May 8;284(19):12585-92
references.pmid: 19129182
references.citation: J Biol Chem. 2004 Aug 20;279(34):35219-27
references.pmid: 15210711
references.citation: Science. 2009 Aug 28;325(5944):1089-93
references.pmid: 19713519
references.citation: J Biol Chem. 2007 Oct 5;282(40):29521-30
references.pmid: 17690105
references.citation: Mol Microbiol. 2002 Apr;44(2):431-48
references.pmid: 11972781
references.citation: Mol Microbiol. 2006 Aug;61(4):847-60
references.pmid: 16879643
references.citation: Mol Microbiol. 2007 Mar;63(5):1331-44
references.pmid: 17244193
references.citation: Anal Biochem. 1979 Feb;93(1):98-102
references.pmid: 434474
references.citation: Biochemistry. 1982 Dec 21;21(26):6628-33
references.pmid: 7159551
references.citation: J Biol Chem. 2002 Apr 19;277(16):13609-14
references.pmid: 11832487
references.citation: Free Radic Biol Med. 2008 Sep 1;45(5):549-61
references.pmid: 18544350