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Literature summary for 1.8.1.8 extracted from

  • Argyrou, A.; Vetting, M.W.; Blanchard, J.S.
    Characterization of a new member of the flavoprotein disulfide reductase family of enzymes from Mycobacterium tuberculosis (2004), J. Biol. Chem., 279, 52694-52702.
    View publication on PubMed
Search for more literature for 1.8.1.8

Cloned(Commentary)

Cloned (Comment) Organism
gene lpdA, expression in Escherichia coli strain BL21(DE3) as His-tagged protein Mycobacterium tuberculosis

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant His-tagged enzyme LpdA, hanging drop vapour diffusion method at room temperature, 0.003 ml protein solution containing 25 mg/ml protein, 10 mM HEPES, pH 7.5, mixed with 0.002 ml precipitant solution containing 0.1 M trisodium citrate, pH 5.4-5.8, 2.5-5.0% PEG 6000, 3-7 days depending on the pH, crystallization takes longer at higher pH-value, crystals are immersed in 10% PEG 6000, 100 mM MES, pH 5.75, and 25% glycerol, X-ray diffraction structure determination and analysis at 2.8 A resolution Mycobacterium tuberculosis

Inhibitors

Inhibitors Comment Organism Structure
2'-phospho-ADP-ribose inhibition of 2,6-dimethyl-1,4-benzoquinone reduction Mycobacterium tuberculosis
NADP+ inhibition of 2,6-dimethyl-1,4-benzoquinone reduction Mycobacterium tuberculosis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information steady-state kinetics, ping-pong kinetic mechanism with NAD(P)H and 2,6-dimethyl-1,4-benzoquinone or 5-hydroxy-1,4-naphthoquinone Mycobacterium tuberculosis
0.00039
-
 5-hydroxy-1,4-naphthoquinone pH 7.5, 25°C, NADPH Mycobacterium tuberculosis
0.00044
-
 NADPH pH 7.5, 25°C, with 5-hydroxy-1,4-naphthoquinone Mycobacterium tuberculosis
0.00053
-
 NADPH pH 7.5, 25°C, with 2,6-dimethyl-1,4-benzoquinone Mycobacterium tuberculosis
0.0016
-
 2,6-dimethyl-1,4-benzoquinone pH 7.5, 25°C, NADPH Mycobacterium tuberculosis
0.0054
-
 thio-NAD+ pH 7.5, 25°C, with NADH Mycobacterium tuberculosis
0.0055
-
 5-hydroxy-1,4-naphthoquinone pH 7.5, 25°C, NADH Mycobacterium tuberculosis
0.011
-
 2,6-dimethyl-1,4-benzoquinone pH 7.5, 25°C, NADH Mycobacterium tuberculosis
0.018
-
 thio-NAD+ pH 7.5, 25°C, with NADPH Mycobacterium tuberculosis
0.028
-
 NADH pH 7.5, 25°C, with 5-hydroxy-1,4-naphthoquinone Mycobacterium tuberculosis
0.036
-
 NADH pH 7.5, 25°C, with 2,6-dimethyl-1,4-benzoquinone Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
protein disulfide + NAD(P)H Mycobacterium tuberculosis
-
 protein dithiol + NAD(P)+
-
 r

Organism

Organism UniProt Comment Textmining
Mycobacterium tuberculosis
-
 gene lpdA
-

Purification (Commentary)

Purification (Comment) Organism
recombinant His-tagged enzyme from Escherichia coli Mycobacterium tuberculosis

Reaction

Reaction Comment Organism Reaction ID
protein-dithiol + NAD(P)+ = protein-disulfide + NAD(P)H + H+ detailed reaction mechanism Mycobacterium tuberculosis
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2,6-dimethyl-1,4-benzoquinone + NAD(P)H
-
 Mycobacterium tuberculosis 2,6-dimethyl-1,4-benzoquinol + NAD(P)+
-
 ?
5-hydroxy-1,4-naphthoquinone + NAD(P)H
-
 Mycobacterium tuberculosis 5-hydroxy-1,4-naphthoquinol + NAD(P)+
-
 ?
additional information enzyme cannot catalyze the reduction of lipoyl substrates, because it lacks one of two cysteine residues involved in dithiol-disulfide interchange with lipoyl substrates and a His-Glu pair involved in general acid catalysis, no reduction of disulfide-bonded substrates Mycobacterium tuberculosis ?
-
 ?
NAD+ + NADH enzyme-catalyzed interconversion Mycobacterium tuberculosis NADH + NAD+
-
 ?
protein disulfide + NAD(P)H
-
 Mycobacterium tuberculosis protein dithiol + NAD(P)+
-
 r
thio-NAD+ + NAD(P)H
-
 Mycobacterium tuberculosis NAD+ + thio-NAD(P)H
-
 ?

Subunits

Subunits Comment Organism
tetramer
-
 Mycobacterium tuberculosis

Synonyms

Synonyms Comment Organism
LpdA
-
 Mycobacterium tuberculosis
More enzyme is a member of the flavoprotein disulfide reductase family of enzymes Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Mycobacterium tuberculosis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0077
-
 2,6-dimethyl-1,4-benzoquinone pH 7.5, 25°C, NADPH Mycobacterium tuberculosis
0.0077
-
 NADPH pH 7.5, 25°C, with 2,6-dimethyl-1,4-benzoquinone Mycobacterium tuberculosis
0.0079
-
 5-hydroxy-1,4-naphthoquinone pH 7.5, 25°C, NADPH Mycobacterium tuberculosis
0.0079
-
 NADPH pH 7.5, 25°C, with 5-hydroxy-1,4-naphthoquinone Mycobacterium tuberculosis
0.0087
-
 thio-NAD+ pH 7.5, 25°C, NAD(P)H Mycobacterium tuberculosis
0.0087
-
 NAD(P)H pH 7.5, 25°C, with thio-NAD+ Mycobacterium tuberculosis
2.5
-
 5-hydroxy-1,4-naphthoquinone pH 7.5, 25°C, NADH Mycobacterium tuberculosis
2.5
-
 NADH pH 7.5, 25°C, with 5-hydroxy-1,4-naphthoquinone Mycobacterium tuberculosis
2.8
-
 2,6-dimethyl-1,4-benzoquinone pH 7.5, 25°C, NADH Mycobacterium tuberculosis
2.8
-
 NADH pH 7.5, 25°C, with 2,6-dimethyl-1,4-benzoquinone Mycobacterium tuberculosis
3.6
-
 thio-NAD+ pH 7.5, 25°C, NADH Mycobacterium tuberculosis
3.6
-
 NADH pH 7.5, 25°C, with thio-NAD+ Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Mycobacterium tuberculosis

Cofactor

Cofactor Comment Organism Structure
FAD tightly but noncovalentyl bound, 1 molecule per subunit, reaction mechanism Mycobacterium tuberculosis
NADP+ tightly but noncovalentyl bound, 1 molecule per subunit, physiological cofactor Mycobacterium tuberculosis

Ki Value [mM]

Ki Value [mM] Ki Value maximum [mM] Inhibitor Comment Organism Structure
0.000006
-
 NADP+
-
 Mycobacterium tuberculosis
0.0008
-
 2'-phospho-ADP-ribose
-
 Mycobacterium tuberculosis