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: ?

Regulation summary diagram for msrA

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, 344°)
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 , InterPro:IN-FAMILY:IPR028427 , Panther:IN-FAMILY:PTHR10173 , PDB:Structure:1FF3 , PDB:Structure:2GT3 , PDB:Structure:2IEM , Pfam:IN-FAMILY:PF01625

Gene-Reaction Schematic: ?

Gene-Reaction Schematic

Genetic Regulation Schematic: ?

Genetic regulation schematic for msrA

GO Terms:

Biological Process: GO:0006979 - response to oxidative stress Inferred from experiment Inferred by computational analysis [GOA01, St01]
GO:0030091 - protein repair Inferred from experiment Inferred by computational analysis [GOA01, Ezraty04]
GO:0006464 - cellular protein modification process Inferred by computational analysis [GOA06]
GO:0055114 - oxidation-reduction process Inferred by computational analysis [UniProtGOA11, GOA01]
Molecular Function: GO:0008113 - peptide-methionine (S)-S-oxide reductase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Grimaud01]
GO:0016671 - oxidoreductase activity, acting on a sulfur group of donors, disulfide as acceptor Inferred from experiment Inferred by computational analysis [GOA01, Moskovitz95]
GO:0036456 - L-methionine-(S)-S-oxide reductase activity Inferred from experiment [BoschiMuller00]
GO:0016491 - oxidoreductase activity Inferred by computational analysis [UniProtGOA11]
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]

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:

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.

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:

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:

Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
thioredoxin 1
reduced thioredoxin 2
a reduced thioredoxin

Sequence Features

Protein sequence of methionine sulfoxide reductase A with features indicated

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Rahman92a, UniProt11a, BoschiMuller00]
UniProt: Removed.
Chain 2 -> 212
UniProt: Peptide methionine sulfoxide reductase msrA;
Active-Site 52
UniProt: Cysteine sulfenic acid (-SOH) intermediate.
Mutagenesis-Variant 52
[BoschiMuller00, UniProt11a]
UniProt: Loss of activity.
Disulfide-Bond-Site 52, 199
UniProt: Redox-active; alternate; Non-Experimental Qualifier: probable;
Mutagenesis-Variant 87
[BoschiMuller00, UniProt11a]
UniProt: No effect.
Mutagenesis-Variant 199
[BoschiMuller00, UniProt11a]
UniProt: Decrease in activity.
Disulfide-Bond-Site 199, 207
UniProt: Redox-active; alternate; Non-Experimental Qualifier: probable;
Mutagenesis-Variant 207
[BoschiMuller00, UniProt11a]
UniProt: Decrease in activity.

Gene Local Context (not to scale): ?

Gene local context diagram

Transcription Unit:

Transcription-unit diagram


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


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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.

UniProt11a: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

UniProt15: UniProt Consortium (2015). "UniProt version 2015-01 released on 2015-01-16 00:00:00." Database.

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

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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
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