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Literature summary for 7.1.1.1 extracted from

  • Clarke, D.M.; Bragg, P.D.
    Purification and properties of reconstitutively active nicotinamide nucleotide transhydrogenase of Escherichia coli (1985), Eur. J. Biochem., 149, 517-523.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli JM83 cells Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
N,N'-dicyclohexylcarbodiimide NADH protects the enzyme against inhibition by N,N'-dicyclohexylcarbodiimide, while NADP+ , and to a lesser extent NADPH increase the rate of inhibition Escherichia coli
N,N'-Dicylclohexylcarbodiimide NADH protects from inhibition, NADP+ and to a lesser extent NADPH increase the rate of inhibition Escherichia coli

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.023
-
NADPH
-
Escherichia coli
0.029
-
NADPH membrane-bound enzyme Escherichia coli
0.033
-
acetylpyridine adenine dinucleotide
-
Escherichia coli
0.041
-
acetylpyridine adenine dinucleotide membrane-bound enzyme Escherichia coli

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasmic membrane
-
Escherichia coli
-
-

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
47000
-
1 * 50000 + 1 * 47000, SDS-PAGE Escherichia coli
50000
-
1 * 50000 + 1 * 47000, SDS-PAGE Escherichia coli

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
NADPH + NAD+ + H+[side 1] Escherichia coli
-
NADP+ + NADH + H+[side 2]
-
r
NADPH + NAD+ + H+[side 1] Escherichia coli W-6
-
NADP+ + NADH + H+[side 2]
-
r

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Escherichia coli W-6
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
Escherichia coli
the recombinant enzyme is purified by pre-extraction of the membranes with sodium cholate and Triton X-100, solubilization of the enzyme with sodium deoxycholate in the presence of 1 M potassium chloride, and centrifugation through a 1.1 M sucrose solution. The wild type enzyme is purified by phenyl Sepharose column chromatography, DEAE-Bio-Gel A column chromatography, and NAD+ agarose column chromatography Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0013
-
unpurified native enzyme, at pH 7.5, temperature not specified in the publication Escherichia coli
0.0052
-
unpurified recombinant enzyme, at pH 7.5, temperature not specified in the publication Escherichia coli
0.022
-
purified native enzyme, at pH 7.5, temperature not specified in the publication Escherichia coli
0.0299
-
purified recombinant enzyme, at pH 7.5, temperature not specified in the publication Escherichia coli
22
-
partially purified enzyme from strain W6 Escherichia coli
29.9
-
purified enzyme from strain JM83 Escherichia coli

Storage Stability

Storage Stability Organism
4°C, 50 mM Tris-HCl, pH 7.8, 1 mM, EDTA, 1 mM, dithiothreitol, 1 week, 10% loss of activity Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
NADPH + 3-acetylpyridine-NAD+
-
Escherichia coli 3-acetylpyridine-NADH + NADP+
-
?
NADPH + 3-acetylpyridine-NAD+
-
Escherichia coli W-6 3-acetylpyridine-NADH + NADP+
-
?
NADPH + NAD+ + H+[side 1]
-
Escherichia coli NADP+ + NADH + H+[side 2]
-
r
NADPH + NAD+ + H+[side 1]
-
Escherichia coli W-6 NADP+ + NADH + H+[side 2]
-
r
NADPH + oxidized acetyl pyridine adenine dinucleotide + H+[side 1]
-
Escherichia coli NADP+ + reduced acetyl pyridine adenine dinucleotide + H+[side 2]
-
r
NADPH + oxidized acetyl pyridine adenine dinucleotide + H+[side 1]
-
Escherichia coli W-6 NADP+ + reduced acetyl pyridine adenine dinucleotide + H+[side 2]
-
r

Subunits

Subunits Comment Organism
heterodimer 1 * 50000 + 1 * 47000, SDS-PAGE Escherichia coli

Synonyms

Synonyms Comment Organism
nicotinamide nucleotide transhydrogenase
-
Escherichia coli

Cofactor

Cofactor Comment Organism Structure
NAD+
-
Escherichia coli
NADH
-
Escherichia coli
NADP+
-
Escherichia coli
NADPH
-
Escherichia coli