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Escherichia coli K-12 substr. MG1655 Enzyme: methionine sulfoxide reductase A



Gene: msrA Accession Numbers: EG11433 (EcoCyc), b4219, ECK4215

Synonyms: pms, pmsR

Regulation Summary Diagram: ?

Summary:
MsrA is one of several methionine sulfoxide reductases in E. coli [Etienne03, Ezraty05]. Methionine residues in proteins can be damaged by oxidation and can be repaired by methionine sulfoxide reductases [Ezraty04]. Conversely, the transcription factor HypT is activated by methionine oxidation and inactivated by MsrA/MsrB activity [Drazic13].

A methionine auxotroph can utilize methionine sulfoxide for growth [Ejiri79, Russel86].

MsrA appears to use thioredoxin 1 as the electron donor in vivo [Russel86, BoschiMuller01, Jacob11]. Overexpressed thioredoxin 2 can substitute for Trx1 [Jacob11].

Cys51, Cys198, and Cys206 are involved in catalysis. A three-step ping-pong reaction mechanism involving a sulfenic acid intermediate on Cys51 has been proposed [BoschiMuller00, BoschiMuller01]. The N and C termini of MsrA are not essential for reductase activity (with DTT), though the C terminus is required for thioredoxin-dependent reduction, and a mutant protein lacking both termini displays kinetic defects [BoschiMuller01]. The rate-limiting step is associated with recycling of thioredoxin [Kriznik14]. A crystal structure is presented at 1.9 Å resolution [TeteFavier00, TeteFavier00a]. Solution structures of both the reduced and oxidized forms of MsrA provide further insight into the catalytic mechanism [Coudevylle07].

Some organisms produce a methionine sulfoxide reductase composed of two domains with similarity to MsrA and MsrB, respectively, whereas E. coli produces these two distinct polypeptides. MsrB exhibits 1000-fold lower catalytic efficiency than MsrA toward free methionine sulfoxide [Grimaud01]. MsrA and MsrB exhibit similar activity (quantitatively) toward an oxidized peptide substrate, though they appear to exhibit some differences in specificity for substrate sites within the peptide [Grimaud01, Ezraty04]. The E. coli MsrA and MsrB polypeptides do not interact with each other in vitro or in a yeast two-hybrid test [Grimaud01].

MsrA appears to have some specificity for the S enantiomer of methionine sulfoxide [Moskovitz00, Etienne03]. Additional methionine sulfoxide reductase activities have been observed in a msrA msrB double mutant, including activities toward both the met-R-(o) and met-S-(o) enantiomers of free (non-peptide) methionine sulfoxide (fRMsr and fSMsr activities) [Etienne03], a soluble NADPH-dependent activity toward the met-S-(o) enantiomer of peptide-linked methionine sulfoxide (MsrA1 activity) [Etienne03, Spector03], and a membrane-associated NADPH-dependent activity toward both met-S-(o) and met-R-(o) enantiomers of peptide-linked and free methionine sulfoxide [Etienne03, Spector03].

Production of MsrA protein is regulated by growth phase [Moskovitz95]. An msrA mutant shows increased sensitivity to oxidative stress [Moskovitz95, St01] as well as increased sensitivity to nitrite and S-nitrosoglutathione, which are released by activated macrophages during infection [St01]. Streptococcus pneumoniae, Neisseria gonorrhoeae, and E. coli (strain MC1061) MsrA proteins are involved in host cell adhesion [Wizemann96].

MsrA: "methionine sulfoxide reductase A" [Rahman94]

Reviews: [Weissbach05, Ezraty05a, BoschiMuller08]

Citations: [Rahman92, Brot81, Levine99, Lin99, Alexander00, Vaughan02, Coudevylle04, Coudevylle06]

Locations: cytosol

Map Position: [4,439,561 <- 4,440,199] (95.69 centisomes)
Length: 639 bp / 212 aa

Molecular Weight of Polypeptide: 23.315 kD (from nucleotide sequence), 24.0 kD (experimental) [Rahman92a ]

Unification Links: ASAP:ABE-0013804 , CGSC:34400 , EchoBASE:EB1403 , EcoGene:EG11433 , EcoliWiki:b4219 , OU-Microarray:b4219 , PortEco:msrA , PR:PRO_000023298 , Pride:P0A744 , Protein Model Portal:P0A744 , RefSeq:NP_418640 , RegulonDB:EG11433 , SMR:P0A744 , String:511145.b4219 , UniProt:P0A744

Relationship Links: InterPro:IN-FAMILY:IPR002569 , PDB:Structure:1FF3 , PDB:Structure:2GT3 , PDB:Structure:2IEM , Pfam:IN-FAMILY:PF01625

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006979 - response to oxidative stress Inferred from experiment Inferred by computational analysis [GOA01a, St01]
GO:0030091 - protein repair Inferred from experiment Inferred by computational analysis [GOA01a, Ezraty04]
GO:0006464 - cellular protein modification process Inferred by computational analysis [GOA06]
GO:0055114 - oxidation-reduction process Inferred by computational analysis [UniProtGOA11a, GOA01a]
Molecular Function: GO:0008113 - peptide-methionine (S)-S-oxide reductase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, GOA01a, Grimaud01]
GO:0016671 - oxidoreductase activity, acting on a sulfur group of donors, disulfide as acceptor Inferred from experiment Inferred by computational analysis [GOA01a, Moskovitz95]
GO:0036456 - L-methionine-(S)-S-oxide reductase activity Inferred from experiment [BoschiMuller00]
GO:0016491 - oxidoreductase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment [LopezCampistrou05, Watt07]

MultiFun Terms: cell processes adaptations other (mechanical, nutritional, oxidative stress)
cell processes defense/survival
information transfer protein related chaperoning, repair (refolding)

Essentiality data for msrA knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Credits:
Last-Curated ? 02-Jul-2014 by Keseler I , SRI International


Enzymatic reaction of: protein-methionine-S-oxide reductase (methionine sulfoxide reductase A)

Synonyms: methionine sulfoxide (protein) reductase, peptide methionine sulfoxide reductase, Met(O)-peptide reductase, protein (methionine sulfoxide) reductase, methionine sulfoxide peptide reductase, protein-L-methionine:oxidized-thioredoxin S-oxidoreductase

EC Number: 1.8.4.11

a protein-L-methionine-(S)-S-oxide + a reduced thioredoxin <=> a [protein]-L-methionine + an oxidized thioredoxin + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

Summary:
The purification and cloning reported in [Rahman92a] was performed with E. coli B. The protein sequences of the K-12 and B enzymes are 100% identical.


Enzymatic reaction of: methionine sulfoxide reductase

Synonyms: methyl sulfoxide reductase, methionine-S-oxide reductase, acetylmethionine sulfoxide reductase

EC Number: 1.8.4.13

L-methionine-(S)-S-oxide + a reduced thioredoxin <=> L-methionine + an oxidized thioredoxin + H2O

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

Alternative Substrates for L-methionine-(S)-S-oxide: Sulindac [Etienne03a ]

Alternative Substrates for a reduced thioredoxin: reduced thioredoxin 2 [Jacob11 ] , thioredoxin 1 [Jacob11 ] , dithiothreitol [BoschiMuller00 ]

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
thioredoxin 1
6.0
3.1
[Jacob11]
L-methionine-(S)-S-oxide
170.0
0.33
[Grimaud01]
reduced thioredoxin 2
37.0
2.7
[Jacob11]
a reduced thioredoxin
10.0
3.7
[BoschiMuller01]


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Rahman92a, UniProt11, BoschiMuller00]
UniProt: Removed.
Chain 2 -> 212
[UniProt09]
UniProt: Peptide methionine sulfoxide reductase msrA;
Mutagenesis-Variant 52
[BoschiMuller00, UniProt11]
Alternate sequence: C → S; UniProt: Loss of activity.
Active-Site 52
[UniProt10a]
UniProt: Cysteine sulfenic acid (-SOH) intermediate;
Disulfide-Bond-Site 199, 52
[UniProt10]
UniProt: Redox-active; alternate; Non-Experimental Qualifier: probable;
Mutagenesis-Variant 87
[BoschiMuller00, UniProt11]
Alternate sequence: C → S; UniProt: No effect.
Mutagenesis-Variant 199
[BoschiMuller00, UniProt11]
Alternate sequence: C → S; UniProt: Decrease in activity.
Disulfide-Bond-Site 207, 199
[UniProt10]
UniProt: Redox-active; alternate; Non-Experimental Qualifier: probable;
Mutagenesis-Variant 207
[BoschiMuller00, UniProt11]
Alternate sequence: C → S; UniProt: Decrease in activity.


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b4219 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG11433; confirmed by SwissProt match.


References

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BoschiMuller00: Boschi-Muller S, Azza S, Sanglier-Cianferani S, Talfournier F, Van Dorsselear A, Branlant G (2000). "A sulfenic acid enzyme intermediate is involved in the catalytic mechanism of peptide methionine sulfoxide reductase from Escherichia coli." J Biol Chem 275(46);35908-13. PMID: 10964927

BoschiMuller01: Boschi-Muller S, Azza S, Branlant G (2001). "E. coli methionine sulfoxide reductase with a truncated N terminus or C terminus, or both, retains the ability to reduce methionine sulfoxide." Protein Sci 10(11);2272-9. PMID: 11604533

BoschiMuller08: Boschi-Muller S, Gand A, Branlant G (2008). "The methionine sulfoxide reductases: Catalysis and substrate specificities." Arch Biochem Biophys 474(2);266-73. PMID: 18302927

Brot81: Brot N, Weissbach L, Werth J, Weissbach H (1981). "Enzymatic reduction of protein-bound methionine sulfoxide." Proc Natl Acad Sci U S A 78(4);2155-8. PMID: 7017726

Coudevylle04: Coudevylle N, Thureau A, Azza S, Boshi-Muller S, Branlant G, Cung MT (2004). "(1)H, (13)C and (15)N resonance assignment of the reduced form of methionine sulfoxide reductase A from Escherichia coli." J Biomol NMR 30(3);363-4. PMID: 15756465

Coudevylle06: Coudevylle N, Antoine M, Boschi-Muller S, Branlant G, Cung MT (2006). "1H, 13C and 15N resonance assignment of an oxidized form (Cys51-Cys198) of methionine sulfoxide reductase A from Escherichia coli." J Biomol NMR 36 Suppl 1;19. PMID: 16505957

Coudevylle07: Coudevylle N, Antoine M, Bouguet-Bonnet S, Mutzenhardt P, Boschi-Muller S, Branlant G, Cung MT (2007). "Solution structure and backbone dynamics of the reduced form and an oxidized form of E. coli methionine sulfoxide reductase A (MsrA): structural insight of the MsrA catalytic cycle." J Mol Biol 366(1);193-206. PMID: 17157315

Drazic13: Drazic A, Miura H, Peschek J, Le Y, Bach NC, Kriehuber T, Winter J (2013). "Methionine oxidation activates a transcription factor in response to oxidative stress." Proc Natl Acad Sci U S A 110(23);9493-8. PMID: 23690622

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Ejiri79: Ejiri SI, Weissbach H, Brot N (1979). "Reduction of methionine sulfoxide to methionine by Escherichia coli." J Bacteriol 139(1);161-4. PMID: 37234

Eklund84: Eklund H, Cambillau C, Sjoberg BM, Holmgren A, Jornvall H, Hoog JO, Branden CI (1984). "Conformational and functional similarities between glutaredoxin and thioredoxins." EMBO J 1984;3(7);1443-9. PMID: 6378624

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Ezraty04: Ezraty B, Grimaud R, El Hassouni M, Moinier D, Barras F (2004). "Methionine sulfoxide reductases protect Ffh from oxidative damages in Escherichia coli." EMBO J 23(8);1868-77. PMID: 15057280

Ezraty05: Ezraty B, Bos J, Barras F, Aussel L (2005). "Methionine sulfoxide reduction and assimilation in Escherichia coli: new role for the biotin sulfoxide reductase BisC." J Bacteriol 187(1);231-7. PMID: 15601707

Ezraty05a: Ezraty B, Aussel L, Barras F (2005). "Methionine sulfoxide reductases in prokaryotes." Biochim Biophys Acta 1703(2);221-9. PMID: 15680230

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

Gleason88: Gleason FK, Holmgren A (1988). "Thioredoxin and related proteins in procaryotes." FEMS Microbiol Rev 1988;4(4);271-97. PMID: 3152490

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GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

Grimaud01: Grimaud R, Ezraty B, Mitchell JK, Lafitte D, Briand C, Derrick PJ, Barras F (2001). "Repair of oxidized proteins. Identification of a new methionine sulfoxide reductase." J Biol Chem 276(52);48915-20. PMID: 11677230

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Jacob11: Jacob C, Kriznik A, Boschi-Muller S, Branlant G (2011). "Thioredoxin 2 from Escherichia coli is not involved in vivo in the recycling process of methionine sulfoxide reductase activities." FEBS Lett 585(12);1905-9. PMID: 21570393

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

Katti90: Katti SK, LeMaster DM, Eklund H (1990). "Crystal structure of thioredoxin from Escherichia coli at 1.68 A resolution." J Mol Biol 1990;212(1);167-84. PMID: 2181145

Kriznik14: Kriznik A, Boschi-Muller S, Branlant G (2014). "Kinetic evidence that methionine sulfoxide reductase A can reveal its oxidase activity in the presence of thioredoxin." Arch Biochem Biophys 548;54-9. PMID: 24632144

Levine99: Levine RL, Berlett BS, Moskovitz J, Mosoni L, Stadtman ER (1999). "Methionine residues may protect proteins from critical oxidative damage." Mech Ageing Dev 107(3);323-32. PMID: 10360685

Lin99: Lin TY (1999). "G33D mutant thioredoxin primarily affects the kinetics of reaction with thioredoxin reductase. Probing the structure of the mutant protein." Biochemistry 38(47);15508-13. PMID: 10569933

LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

Moskovitz00: Moskovitz J, Poston JM, Berlett BS, Nosworthy NJ, Szczepanowski R, Stadtman ER (2000). "Identification and characterization of a putative active site for peptide methionine sulfoxide reductase (MsrA) and its substrate stereospecificity." J Biol Chem 275(19);14167-72. PMID: 10799493

Moskovitz95: Moskovitz J, Rahman MA, Strassman J, Yancey SO, Kushner SR, Brot N, Weissbach H (1995). "Escherichia coli peptide methionine sulfoxide reductase gene: regulation of expression and role in protecting against oxidative damage." J Bacteriol 177(3);502-7. PMID: 7836279

Nikkola93: Nikkola M, Gleason FK, Fuchs JA, Eklund H (1993). "Crystal structure analysis of a mutant Escherichia coli thioredoxin in which lysine 36 is replaced by glutamic acid." Biochemistry 1993;32(19);5093-8. PMID: 8098620

Rahman92: Rahman MA, Brot N, Weissbach H (1992). "High level expression and purification of peptide methionine sulfoxide reductase in Escherichia coli." Cell Mol Biol 38(5);529-42. PMID: 1468111

Rahman92a: Rahman MA, Nelson H, Weissbach H, Brot N (1992). "Cloning, sequencing, and expression of the Escherichia coli peptide methionine sulfoxide reductase gene." J Biol Chem 267(22);15549-51. PMID: 1386361

Rahman94: Rahman MA, Moskovitz J, Strassman J, Weissbach H, Brot N (1994). "Physical map location of the peptide methionine sulfoxide reductase gene on the Escherichia coli chromosome." J Bacteriol 176(5);1548-9. PMID: 8113203

Russel86: Russel M, Model P (1986). "The role of thioredoxin in filamentous phage assembly. Construction, isolation, and characterization of mutant thioredoxins." J Biol Chem 261(32);14997-5005. PMID: 3533930

Spector03: Spector D, Etienne F, Brot N, Weissbach H (2003). "New membrane-associated and soluble peptide methionine sulfoxide reductases in Escherichia coli." Biochem Biophys Res Commun 302(2);284-9. PMID: 12604343

St01: St John G, Brot N, Ruan J, Erdjument-Bromage H, Tempst P, Weissbach H, Nathan C (2001). "Peptide methionine sulfoxide reductase from Escherichia coli and Mycobacterium tuberculosis protects bacteria against oxidative damage from reactive nitrogen intermediates." Proc Natl Acad Sci U S A 98(17);9901-6. PMID: 11481433

TeteFavier00: Tete-Favier F, Cobessi D, Leonard GA, Azza S, Talfournier F, Boschi-Muller S, Branlant G, Aubry A (2000). "Crystallization and preliminary X-ray diffraction studies of the peptide methionine sulfoxide reductase from Escherichia coli." Acta Crystallogr D Biol Crystallogr 56(Pt 9);1194-7. PMID: 10957644

TeteFavier00a: Tete-Favier F, Cobessi D, Boschi-Muller S, Azza S, Branlant G, Aubry A (2000). "Crystal structure of the Escherichia coli peptide methionine sulphoxide reductase at 1.9 A resolution." Structure 8(11);1167-78. PMID: 11080639

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

Vaughan02: Vaughan MD, Sampson PB, Honek JF (2002). "Methionine in and out of proteins: targets for drug design." Curr Med Chem 9(3);385-409. PMID: 11860363

Watt07: Watt RM, Wang J, Leong M, Kung HF, Cheah KS, Liu D, Danchin A, Huang JD (2007). "Visualizing the proteome of Escherichia coli: an efficient and versatile method for labeling chromosomal coding DNA sequences (CDSs) with fluorescent protein genes." Nucleic Acids Res 35(6);e37. PMID: 17272300

Weissbach05: Weissbach H, Resnick L, Brot N (2005). "Methionine sulfoxide reductases: history and cellular role in protecting against oxidative damage." Biochim Biophys Acta 1703(2);203-12. PMID: 15680228

Wizemann96: Wizemann TM, Moskovitz J, Pearce BJ, Cundell D, Arvidson CG, So M, Weissbach H, Brot N, Masure HR (1996). "Peptide methionine sulfoxide reductase contributes to the maintenance of adhesins in three major pathogens." Proc Natl Acad Sci U S A 93(15);7985-90. PMID: 8755589

Other References Related to Gene Regulation

Bradley07: Bradley MD, Beach MB, de Koning AP, Pratt TS, Osuna R (2007). "Effects of Fis on Escherichia coli gene expression during different growth stages." Microbiology 153(Pt 9);2922-40. PMID: 17768236


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Fri Dec 19, 2014, biocyc14.