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Literature summary extracted from
- Purification and characterization of a rotenone-insensitive NADH:Q6 oxidoreductase from mitochondria of Saccharomyces cerevisiae (1988), Eur. J. Biochem., 176, 377-384.
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.6.5.9 | flavone | - |
Saccharomyces cerevisiae |  |
| 1.6.5.9 | additional information | NAD+, Mg-ADP, Mg-ATP, Ca2+ (all at 2 mM) or piericidin, rotenone (both at 0.01 mM) or adenosyl-3'-O-[3-[N-(4-azido-2-nitrophenyl)-amino]propionyl]-NAD+ (0.05 mM) are without inhibitory or stimulatory effects | Saccharomyces cerevisiae |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 7.1.1.2 | 0.031 | - |
NADH | with ubiquinone-6 as acceptor | Saccharomyces cerevisiae | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 1.6.5.9 | mitochondrial membrane | mitochondrial inner membrane | Saccharomyces cerevisiae | 31966 | - |
| 7.1.1.2 | mitochondrion | - |
Saccharomyces cerevisiae | 5739 | - |
Metals/Ions
| EC Number | Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|---|
| 7.1.1.2 | Iron | contains 0.08 mol Fe per mol of flavin after dialysis | Saccharomyces cerevisiae | |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 1.6.5.9 | 53000 | - |
SDS-PAGE | Saccharomyces cerevisiae |
| 7.1.1.2 | 53000 | - |
x * 53000, SDS-PAGE | Saccharomyces cerevisiae |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 1.6.5.9 | Saccharomyces cerevisiae | - |
- |
- |
| 7.1.1.2 | Saccharomyces cerevisiae | - |
rotenone-insensitive enzyme | - |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 1.6.5.9 | DEAE-cellulose column chromatography and blue Sepharose CL-6B column chromatography | Saccharomyces cerevisiae |
| 7.1.1.2 | - |
Saccharomyces cerevisiae |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 1.6.5.9 | 0.78 | - |
crude extract, using ubiquinone-2 as substrate, at pH 6.2, temperature not specified in the publication | Saccharomyces cerevisiae |
| 1.6.5.9 | 61.9 | - |
after 2134fold purification, using ubiquinone-6 as substrate, at pH 6.2, temperature not specified in the publication | Saccharomyces cerevisiae |
| 1.6.5.9 | 1671 | - |
after 2134fold purification, using ubiquinone-2 as substrate, at pH 6.2, temperature not specified in the publication | Saccharomyces cerevisiae |
| 7.1.1.2 | 61.9 | - |
reaction with ubiquinone-6 | Saccharomyces cerevisiae |
| 7.1.1.2 | 1671 | - |
reaction with ubiquinone-2 | Saccharomyces cerevisiae |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 1.6.5.9 | 2 ferricyanide + NADH | - |
Saccharomyces cerevisiae | 2 ferrocyanide + NAD+ + H+ | - |
? | |
| 1.6.5.9 | NADH + H+ + 2,6-dichloroindophenol | - |
Saccharomyces cerevisiae | NAD+ + reduced 2,6-dichloroindophenol | - |
? | |
| 1.6.5.9 | NADH + H+ + menaquinone | - |
Saccharomyces cerevisiae | NAD+ + menaquinol | - |
? | |
| 1.6.5.9 | NADH + H+ + ubiquinone-10 | - |
Saccharomyces cerevisiae | NAD+ + ubiquinol-10 | - |
? | |
| 1.6.5.9 | NADH + H+ + ubiquinone-2 | ubiquinone is the best electron acceptor | Saccharomyces cerevisiae | NAD+ + ubiquinol-2 | - |
? | |
| 1.6.5.9 | NADH + H+ + ubiquinone-6 | - |
Saccharomyces cerevisiae | NAD+ + ubiquinol-6 | - |
? | |
| 7.1.1.2 | NADH + ubiquinone | - |
Saccharomyces cerevisiae | NAD+ + ubiquinol | - |
? | |
| 7.1.1.2 | NADH + ubiquinone-2 | - |
Saccharomyces cerevisiae | NAD+ + reduced ubiquinol-2 | - |
? | |
| 7.1.1.2 | NADH + ubiquinone-6 | - |
Saccharomyces cerevisiae | NAD+ + ubiquinol-6 | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 1.6.5.9 | monomer | 1 * 53000, SDS-PAGE | Saccharomyces cerevisiae |
| 7.1.1.2 | ? | x * 53000, SDS-PAGE | Saccharomyces cerevisiae |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 1.6.5.9 | NADH:Q6 oxidoreductase | - |
Saccharomyces cerevisiae |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 1.6.5.9 | 0.031 | - |
NADH | using ubiquinone-2 as substrate, in 20 mM MOPS/KOH, 1 mM EDTA, 0.03% (v/v) Triton X-100, 200 mM KCl, pH 6.2, temperature not specified in the publication | Saccharomyces cerevisiae | |
| 7.1.1.2 | 80 | 100 | NADH | with ubiquinone-6 as acceptor, Triton assay medium | Saccharomyces cerevisiae | |
| 7.1.1.2 | 500 | 550 | NADH | with ubiquinone-6 as acceptor, mitochondria | Saccharomyces cerevisiae | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.6.5.9 | 6.2 | - |
with ubiquinone-2 as acceptor a sharp maximum at pH 6.2 is obtained | Saccharomyces cerevisiae |
| 7.1.1.2 | 4.5 | 9.5 | reaction with ubiquinone-6 is almost constant between pH 4.5 and pH 9.5 | Saccharomyces cerevisiae |
| 7.1.1.2 | 6.2 | - |
reaction with ubiquinone-2 | Saccharomyces cerevisiae |
pH Range
| EC Number | pH Minimum | pH Maximum | Comment | Organism |
|---|---|---|---|---|
| 1.6.5.9 | 4.5 | 9.5 | with ubiquinone-6 as acceptor the activity is almost constant between pH 4.5 and 9.5, at pH values lower than 4.5 and higher than 9.5, enzymic activity is irreversibly destroyed | Saccharomyces cerevisiae |
pH Stability
| EC Number | pH Stability | pH Stability Maximum | Comment | Organism |
|---|---|---|---|---|
| 7.1.1.2 | 4.5 | - |
activity is irreversibly destroyed below | Saccharomyces cerevisiae |
| 7.1.1.2 | 9.5 | - |
activity is irreversibly destroyed above | Saccharomyces cerevisiae |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 1.6.5.9 | FAD | non-covalently linked | Saccharomyces cerevisiae | |
| 1.6.5.9 | additional information | the enzyme contains no iron-sulfur clusters | Saccharomyces cerevisiae | |
| 1.6.5.9 | NADH | the enzyme is specific for NADH. NADPH or deamino-NADH show rates that are at least 250times lower than obtained with NADH at any pH between 5.5 and 8.5 | Saccharomyces cerevisiae | |
| 7.1.1.2 | FAD | contains a single molecule of FAD per subunit | Saccharomyces cerevisiae | |
| 7.1.1.2 | NADH | - |
Saccharomyces cerevisiae | |
IC50 Value
| EC Number | IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
|---|---|---|---|---|---|---|
| 1.6.5.9 | 0.095 | - |
using ubiquinone-2 as substrate, in 20 mM MOPS/KOH, 1 mM EDTA, 0.03% (v/v) Triton X-100, 200 mM KCl, pH 6.2, temperature not specified in the publication | Saccharomyces cerevisiae | flavone | |
Expression
| EC Number | Organism | Comment | Expression |
|---|---|---|---|
| 1.6.5.9 | Saccharomyces cerevisiae | the amount of enzyme in the cell is subject to glucose repression, it increases slightly when cells, grown on glucose or lactate, enter the stationary phase | up |
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