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Literature summary for 1.8.4.12 extracted from

  • Lowther, W.T.; Weissbach, H.; Etienne, F.; Brot, N.; Matthews, B.W.
    The mirrored methionine sulfoxide reductases of Neisseria gonorrhoeae pilB (2002), Nat. Struct. Biol., 9, 348-352.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
CBS-1, overexpression of the His-tagged enzyme in Escherichia coli Homo sapiens
overexpression of the full length tandem enzyme, the MsrA, and the MsrB domains, all His-tagged, in Escherichia coli Neisseria gonorrhoeae
yeaA, overexpression of the His-tagged enzyme in Escherichia coli Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant detagged MsrB domain containing SeMet39, hanging drop vapour-diffusion method, 15 mg/ml protein in 20 mM Tris, pH 8.5, 10% v/v glycerol, against a well solution containing 0.1 M sodium cacodylate, pH 6.5, 30% w/v PEG 4000, room temperature, X-ray diffraction structure determination and analysis at 1.8 A resolution, modeling Neisseria gonorrhoeae

Organism

Organism UniProt Comment Textmining
Escherichia coli P0A746
-
-
Homo sapiens Q9Y3D2
-
-
Neisseria gonorrhoeae P14930 enzyme contains MsrA and MsrB domains; PilB enzyme has 2 catalytic domains showing MsrA, methionine S-oxide reductase (S-form oxidizing), and MsrB, methionine S-oxide reductase (R-form oxidizing), activity, respectively, the MsrB domain contains a selenomethionine at position 39
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli Homo sapiens
recombinant His-tagged enzyme from Escherichia coli Escherichia coli
recombinant His-tagged full length tandem enzyme, MsrA, and MsrB domains from Escherichia coli, tags are removed by thrombin digestion Neisseria gonorrhoeae

Reaction

Reaction Comment Organism Reaction ID
L-methionine (R)-sulfoxide + thioredoxin = L-methionine + thioredoxin disulfide + H2O reaction mechanism, modeling of substrate binding at the active site, Cys444 and Cys495 are involved Neisseria gonorrhoeae

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.075
-
purified recombinant MsrA/MsrB tandem domain, substrate L-methionine (R)-sulfoxide Neisseria gonorrhoeae
0.085
-
purified recombinant CBS-1, substrate L-methionine (R)-sulfoxide Escherichia coli
0.092
-
purified recombinant CBS-1, substrate L-methionine (R)-sulfoxide Homo sapiens
0.11
-
purified recombinant MsrB domain alone, substrate L-methionine (R)-sulfoxide Neisseria gonorrhoeae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-methionine (R)-sulfoxide + thioredoxin MsrB activity of the tandem domains of PilB, the MsrB domain alone does not utilize the S-isomer Neisseria gonorrhoeae L-methionine + thioredoxin disulfide + H2O
-
?
L-methionine (R)-sulfoxide + thioredoxin MsrB is specific for the R-isomer, no activity with the S-isomer Homo sapiens L-methionine + thioredoxin disulfide + H2O
-
?
L-methionine (R)-sulfoxide + thioredoxin MsrB is specific for the R-isomer, no activity with the S-isomer Escherichia coli L-methionine + thioredoxin disulfide + H2O
-
?
additional information the tandem domains of PilB also possess MsrA activity utilizing L-methionine (S)-sulfoxide as substrate, the MsrA domain alone does very poorly utilize the R-isomer Neisseria gonorrhoeae ?
-
?

Subunits

Subunits Comment Organism
More enzyme domain and active site structure, MsrB domain comprises residues 375-522, overview Neisseria gonorrhoeae

Synonyms

Synonyms Comment Organism
CBS-1
-
Homo sapiens
methionine sulfoxide reductase
-
Neisseria gonorrhoeae
methionine sulfoxide reductase
-
Homo sapiens
methionine sulfoxide reductase
-
Escherichia coli
methionine sulfoxide reductase B
-
Neisseria gonorrhoeae
MsrB
-
Neisseria gonorrhoeae
PilB
-
Neisseria gonorrhoeae
YeaA
-
Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
assay at Neisseria gonorrhoeae
37
-
assay at Homo sapiens
37
-
assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.9
-
assay at Neisseria gonorrhoeae
6.9
-
assay at Homo sapiens
6.9
-
assay at Escherichia coli

Cofactor

Cofactor Comment Organism Structure
thioredoxin
-
Neisseria gonorrhoeae
thioredoxin
-
Homo sapiens
thioredoxin
-
Escherichia coli