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Literature summary extracted from

  • Steffek, M.; Newton, G.L.; Av-Gay, Y.; Fahey, R.C.
    Characterization of Mycobacterium tuberculosis mycothiol S-conjugate amidase (2003), Biochemistry, 42, 12067-12076.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.115 expressed in Escherichia coli BL21(DE3) cells Mycobacterium tuberculosis

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.5.1.115 1,10-phenanthroline 0.19% residual activity at 1 mM 1,10-phenanthroline Mycobacterium tuberculosis
3.5.1.115 1,7-phenanthroline slightly more than 50% inhibition at 1 mM 1,7-phenanthroline Mycobacterium tuberculosis
3.5.1.115 mycothiol about 20% residual activity at 2 mM mycothiol Mycobacterium tuberculosis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.115 0.19
-
RifS13 in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 0.44
-
mycothiol-7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 0.45
-
RifS17 in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 0.65
-
mycothiol-cerulenin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 0.69
-
mycothiol-acetophenone in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 1.3
-
mycothiol-N-ethylmaleimide in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 3
-
mycothiol-3-(N-maleimidopropionyl)biocytin Km above 3.0 mM, in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 4.8
-
mycothiol disulfide in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.115 Ca2+ the protein contains 2.6 equivalents of Ca2+ per subunit after purification Mycobacterium tuberculosis
3.5.1.115 Zn2+ the protein contains 1.4 equivalents of Zn2+ per subunit after purification Mycobacterium tuberculosis

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.5.1.115 32697
-
x * 32697, calculated from amino acid sequence Mycobacterium tuberculosis
3.5.1.115 33000
-
x * 33000, SDA-PAGE Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.1.115 a mycothiol S-conjugate + H2O Mycobacterium tuberculosis the enzyme is extremely specific for the mycothiol portion of the mycothiol S-conjugate and relatively nonspecific with respect to the R-moiety an N-acetyl L-cysteine-S-conjugate + 1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
?
3.5.1.115 a mycothiol S-conjugate + H2O Mycobacterium tuberculosis H37Rv the enzyme is extremely specific for the mycothiol portion of the mycothiol S-conjugate and relatively nonspecific with respect to the R-moiety an N-acetyl L-cysteine-S-conjugate + 1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
?

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.115 Mycobacterium tuberculosis
-
-
-
3.5.1.115 Mycobacterium tuberculosis H37Rv
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.115 ammonium sulfate precipitation, hydroxyapatite column chromatography, phenyl Sepharose column chromatography, and Sephadex G-25 gel filtration Mycobacterium tuberculosis

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.5.1.115 0.13
-
crude extract, in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 10.6
-
after 82fold purification, in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.115 a mycothiol S-conjugate + H2O the enzyme is extremely specific for the mycothiol portion of the mycothiol S-conjugate and relatively nonspecific with respect to the R-moiety Mycobacterium tuberculosis an N-acetyl L-cysteine-S-conjugate + 1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
?
3.5.1.115 a mycothiol S-conjugate + H2O the enzyme is extremely specific for the mycothiol portion of the mycothiol S-conjugate and relatively nonspecific with respect to the R-moiety Mycobacterium tuberculosis H37Rv an N-acetyl L-cysteine-S-conjugate + 1-O-(2-amino-2-deoxy-alpha-D-glucopyranosyl)-1D-myo-inositol
-
?
3.5.1.115 additional information the enzyme is highly specific for the mycothiol moiety of mycothiol S-conjugates and relatively nonspecific for the structure of the sulfur-linked conjugate Mycobacterium tuberculosis ?
-
?
3.5.1.115 additional information the enzyme is highly specific for the mycothiol moiety of mycothiol S-conjugates and relatively nonspecific for the structure of the sulfur-linked conjugate Mycobacterium tuberculosis H37Rv ?
-
?
3.5.1.115 mycothiol disulfide + H2O
-
Mycobacterium tuberculosis ?
-
?
3.5.1.115 mycothiol disulfide + H2O
-
Mycobacterium tuberculosis H37Rv ?
-
?
3.5.1.115 mycothiol-3-(N-maleimidopropionyl)biocytin + H2O
-
Mycobacterium tuberculosis ?
-
?
3.5.1.115 mycothiol-3-(N-maleimidopropionyl)biocytin + H2O
-
Mycobacterium tuberculosis H37Rv ?
-
?
3.5.1.115 mycothiol-7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin + H2O
-
Mycobacterium tuberculosis ?
-
?
3.5.1.115 mycothiol-7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin + H2O
-
Mycobacterium tuberculosis H37Rv ?
-
?
3.5.1.115 mycothiol-acetophenone + H2O
-
Mycobacterium tuberculosis ?
-
?
3.5.1.115 mycothiol-cerulenin + H2O
-
Mycobacterium tuberculosis ?
-
?
3.5.1.115 mycothiol-N-ethylmaleimide + H2O
-
Mycobacterium tuberculosis ?
-
?
3.5.1.115 RifS13 + H2O
-
Mycobacterium tuberculosis 1-D-myo-inositol-alpha-D-glucopyranoside + N-acetyl-S-[(2R,12Z,14E,16S,18S,19S,20S,21S,22S,23S,24E)-5,6,9,17,19-pentahydroxy-23-methoxy-21-(methoxycarbonyl)-2,4,12,16,18,20,22-heptamethyl-1,11-dioxo-1,2-dihydro-2,7-(epoxypentadeca[1,11,13]trienoimino)naphtho[2,1-b]furan-8-yl]-L-cysteine
-
?
3.5.1.115 RifS17 + H2O
-
Mycobacterium tuberculosis 1-D-myo-inositol-alpha-D-glucopyranoside + N-acetyl-S-[(2R,12Z,14E,16S,18S,19S,20S,21S,22S,23S,24E)-5,17,19-trihydroxy-23-methoxy-21-(methoxycarbonyl)-2,4,12,16,18,20,22-heptamethyl-1,6,9,11-tetraoxo-1,2,6,9-tetrahydro-2,7-(epoxypentadeca[1,11,13]trienoimino)naphtho[2,1-b]furan-8-yl]-L-cysteine
-
?

Subunits

EC Number Subunits Comment Organism
3.5.1.115 ? x * 32697, calculated from amino acid sequence Mycobacterium tuberculosis
3.5.1.115 ? x * 33000, SDA-PAGE Mycobacterium tuberculosis

Synonyms

EC Number Synonyms Comment Organism
3.5.1.115 MCA
-
Mycobacterium tuberculosis
3.5.1.115 Rv1082
-
Mycobacterium tuberculosis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.5.1.115 0.82
-
RifS13 in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 1.04
-
RifS17 in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 1.3
-
mycothiol-7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 1.7
-
mycothiol disulfide in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 3
-
mycothiol-3-(N-maleimidopropionyl)biocytin kcat above 3.0 s-1, in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 7
-
mycothiol-cerulenin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 15
-
mycothiol-N-ethylmaleimide in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 37
-
mycothiol-acetophenone in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.5.1.115 0.35
-
mycothiol disulfide in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 1.03
-
mycothiol-3-(N-maleimidopropionyl)biocytin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 2.3
-
RifS17 in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 3
-
mycothiol-7-(diethylamino)-3-(4'-maleimidylphenyl)-4-methylcoumarin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 4.3
-
RifS13 in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 11
-
mycothiol-cerulenin in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 12
-
mycothiol-N-ethylmaleimide in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis
3.5.1.115 54
-
mycothiol-acetophenone in 50 mM HEPES (pH 7.5) containing 50 mM NaCl and 0.1 mM dithiothreitol, at 37°C Mycobacterium tuberculosis