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Literature summary for 1.8.4.12 extracted from

  • Kim, H.Y.; Gladyshev, V.N.
    Different catalytic mechanisms in mammalian selenocysteine- and cysteine-containing methionine-R-sulfoxide reductases (2005), PLoS Biol., 3, 2080-2089.
No PubMed abstract available
Search for more literature for 1.8.4.12

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant isozyme MsrB3 in Escherichia coli strain BL21(DE3) Homo sapiens
expression of wild-type and mutant isozymes MsrB1 and MsrB2 in Escherichia coli strain BL21(DE3) and in NIH-3T3 cells Mus musculus

Protein Variants

Protein Variants Comment Organism
E81V site-directed mutagenesis, mutation in the selenocysteine-containing or the cysteine-containing isozyme MsrB1, both mutants show reduced activity with either cofactor thioredoxin and DTT compared to wild-type MsrB1s Mus musculus
F97N site-directed mutagenesis, mutation in the selenocysteine-containing or the cysteine-containing isozyme MsrB1, both mutants show altered activity and kinetics compared to wild-type MsrB1s Mus musculus
G77H site-directed mutagenesis, mutation in the selenocysteine-containing or the cysteine-containing isozyme MsrB1, the selenocysteine-containing mutant shows reduced activity with either cofactor thioredoxin and DTT compared to wild-type selenocysteine MsrB1, while the cysteine-containing mutant shows activity and kinetics similar to the wild-type cysteine MsrB1 Mus musculus
G77H/E81V/F97N site-directed mutagenesis, mutation in the selenocysteine-containing or the cysteine-containing isozyme MsrB1, both mutants show altered activity and kinetics compared to wild-type MsrB1s Mus musculus
G77H/F97N site-directed mutagenesis, mutation in the selenocysteine-containing or the cysteine-containing isozyme MsrB1, both mutants show altered activity and kinetics compared to wild-type MsrB1s Mus musculus
H77G site-directed mutagenesis, mutation of isozyme MsrB2 leads to highly reduced activity with either cofactor thioredoxin and DTT compared to wild-type MsrB2 Mus musculus
H77G site-directed mutagenesis, mutation of isozyme MsrB3 leads to highly reduced activity with cofactor thioredoxin or DTT compared to wild-type MsrB3 Homo sapiens
H77G/I81E/N97F site-directed mutagenesis, mutation of isozyme MsrB3, inactive mutant Homo sapiens
H77G/N97F site-directed mutagenesis, mutation of isozyme MsrB2, inactive mutant Mus musculus
H77G/N97F site-directed mutagenesis, mutation of isozyme MsrB3, inactive mutant Homo sapiens
H77G/V81E/N97F site-directed mutagenesis, mutation of isozyme MsrB2, inactive mutant Mus musculus
I81E site-directed mutagenesis, mutation of isozyme MsrB3 leads to slightly increased activity with cofactor thioredoxin and reduced activcity with DTT compared to wild-type MsrB3 Homo sapiens
additional information substitution of Cys residues abolish the enzyme's activity with thioredoxin and increase the DTT-dependent activity, overview Mus musculus
additional information substitution of Cys residues abolish the enzyme's activity with thioredoxin and increase the DTT-dependent activity, overview Homo sapiens
N97F site-directed mutagenesis, mutation of isozyme MsrB2, the mutant is inactive with cofactor thioredoxin and shows highly reduced activity with cofactor DTT compared to wild-type MsrB2 Mus musculus
N97F site-directed mutagenesis, mutation of isozyme MsrB3, the mutant is inactive with cofactor thioredoxin and shows highly reduced activity with cofactor DTT compared to wild-type MsrB3 Homo sapiens
N97Y site-directed mutagenesis, mutation of isozyme MsrB2, the mutant is inactive with cofactor thioredoxin and shows highly reduced activity with cofactor DTT compared to wild-type MsrB2 Mus musculus
N97Y site-directed mutagenesis, mutation of isozyme MsrB3, the mutant shows highly reduced activity with cofactor DTT or thioredoxin compared to wild-type MsrB3 Homo sapiens
V81E site-directed mutagenesis, mutation of isozyme MsrB2 leads to reduced activity with either cofactor thioredoxin and DTT compared to wild-type MsrB2 Mus musculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information thioredoxin- and DTT-dependent kinetics of wild-type and mutant enzymes, overview Mus musculus
additional information
-
 additional information thioredoxin- and DTT-dependent kinetics of wild-type and mutant enzymes, overview Homo sapiens
0.31
-
 L-methionine-(R)-S-oxide recombinant wild-type isozyme MsrB2 expressed in Escherichia coli Mus musculus
0.5
-
 L-methionine-(R)-S-oxide recombinant wild-type selenocysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells Mus musculus
0.8
-
 L-methionine-(R)-S-oxide recombinant wild-type isozyme MsrB3 expressed in Escherichia coli Homo sapiens
1.3
-
 L-methionine-(R)-S-oxide recombinant wild-type cysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells Mus musculus

Metals/Ions

Metals/Ions Comment Organism Structure
additional information isozyme MsrB1 contains selenocysteine, while isozymes MsrB2 and MsrB3 contain cysteine residues, the thioredoxin dependence is different for selenocysteine- and cysteine-containing enzyme, overview Mus musculus
additional information native isozyme MsrB1 contains selenocysteine, while native isozymes MsrB2 and MsrB3 contain cysteine residues, the thioredoxin dependence is different for selenocysteine- and cysteine-containing enzyme, overview Homo sapiens

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-methionine-(R)-S-oxide + thioredoxin Mus musculus
-
 L-methionine + thioredoxin disulfide + H2O
-
 ?
L-methionine-(R)-S-oxide + thioredoxin Homo sapiens
-
 L-methionine + thioredoxin disulfide + H2O
-
 ?

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
 isozymes MsrB1, MsrB2, and MsrB3
-
Mus musculus
-
 isozymes MsrB1, MsrB2, and MsrB3
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant isozyme MsrB3 from Escherichia coli strain BL21(DE3) Homo sapiens
recombinant wild-type and mutant isozymes MsrB1 and MsrB2 from Escherichia coli strain BL21(DE3) and NIH-3T3 cells Mus musculus

Reaction

Reaction Comment Organism Reaction ID
peptide-L-methionine + thioredoxin disulfide + H2O = peptide-L-methionine (R)-S-oxide + thioredoxin catalytic mechanism involving residues at positions 95, 41, 97, 77, and 80, molecular modeling, role of selenocysteine- and cysteine residues in catalysis, overview Mus musculus
a
peptide-L-methionine + thioredoxin disulfide + H2O = peptide-L-methionine (R)-S-oxide + thioredoxin catalytic mechanism involving residues at positions 95, 41, 97, 77, and 80, molecular modeling, role of selenocysteine- and cysteine residues in catalysis, overview Homo sapiens
a

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
additional information
-
 thioredoxin- and DTT-dependent activities of mutant enzymes, overview Mus musculus
additional information
-
 thioredoxin- and DTT-dependent activities of mutant enzymes, overview Homo sapiens
0.000046
-
 purified recombinant wild-type cysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells, substrates are L-methionine-(R)-S-oxide and thioredoxin Mus musculus
0.002
-
 purified recombinant wild-type cysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells, substrates are L-methionine-(R)-S-oxide and DTT Mus musculus
0.01
-
 purified recombinant wild-type isozyme MsrB2 expressed in Escherichia coli, substrates are L-methionine-(R)-S-oxide and thioredoxin Mus musculus
0.023
-
 purified recombinant wild-type isozyme MsrB3 expressed in Escherichia coli, substrates are L-methionine-(R)-S-oxide and thioredoxin Homo sapiens
0.045
-
 purified recombinant wild-type selenocysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells, substrates are L-methionine-(R)-S-oxide and thioredoxin Mus musculus
0.17
-
 purified recombinant wild-type selenocysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells, substrates are L-methionine-(R)-S-oxide and DTT Mus musculus
0.386
-
 purified recombinant wild-type isozyme MsrB2 expressed in Escherichia coli, substrates are L-methionine-(R)-S-oxide and DTT Mus musculus
0.452
-
 purified recombinant wild-type isozyme MsrB3 expressed in Escherichia coli, substrates are L-methionine-(R)-S-oxide and DTT Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-methionine-(R)-S-oxide + DTT isozymes MsrB1, MsrB2, and MsrB3 Mus musculus L-methionine + thioredoxin disulfide + H2O
-
 ?
L-methionine-(R)-S-oxide + thioredoxin
-
 Mus musculus L-methionine + thioredoxin disulfide + H2O
-
 ?
L-methionine-(R)-S-oxide + thioredoxin
-
 Homo sapiens L-methionine + thioredoxin disulfide + H2O
-
 ?
L-methionine-(R)-S-oxide + thioredoxin isozymes MsrB1, MsrB2, and MsrB3 Mus musculus L-methionine + thioredoxin disulfide + H2O
-
 ?
L-methionine-(R)-S-oxide + thioredoxin isozymes MsrB1, MsrB2, and MsrB3 Homo sapiens L-methionine + thioredoxin disulfide + H2O
-
 ?
additional information the thioredoxin dependence is different for selenocysteine- and cysteine-containing enzyme, overview Mus musculus ?
-
 ?
additional information the thioredoxin dependence is different for selenocysteine- and cysteine-containing enzyme, overview Homo sapiens ?
-
 ?

Synonyms

Synonyms Comment Organism
cysteine-containing methionine-R-sulfoxide reductase
-
 Mus musculus
cysteine-containing methionine-R-sulfoxide reductase
-
 Homo sapiens
methionine sulfoxide reductase B
-
 Mus musculus
methionine sulfoxide reductase B
-
 Homo sapiens
MsrB
-
 Mus musculus
MsrB
-
 Homo sapiens
selenocysteine-containing methionine-R-sulfoxide reductase
-
 Mus musculus
selenocysteine-containing methionine-R-sulfoxide reductase
-
 Homo sapiens

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.002
-
 L-methionine-(R)-S-oxide recombinant wild-type cysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells Mus musculus
0.22
-
 L-methionine-(R)-S-oxide recombinant wild-type selenocysteine-containing isozyme MsrB1 expressed in NIH 3T3 cells Mus musculus
0.46
-
 L-methionine-(R)-S-oxide recombinant wild-type isozyme MsrB2 expressed in Escherichia coli Mus musculus
0.83
-
 L-methionine-(R)-S-oxide recombinant wild-type isozyme MsrB3 expressed in Escherichia coli Homo sapiens

Cofactor

Cofactor Comment Organism Structure
dithiothreitol results in higher activity compared to cofactor thioredoxin Mus musculus
thioredoxin
-
 Mus musculus