Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant enzymes in strain DW35 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
D95E | site-directed mutagenesis of subunit C, the mutant shows reduced activity and a shifted pH-optimum compared to the wild-type enzyme | Escherichia coli |
D95L | site-directed mutagenesis of subunit C, the mutant shows a shifted pH-optimum but similar activity compared to the wild-type enzyme | Escherichia coli |
E101D | site-directed mutagenesis of subunit C, the mutant shows reduced activity and a shifted pH-optimum compared to the wild-type enzyme | Escherichia coli |
E101L | site-directed mutagenesis of subunit C, the mutant shows reduced activity and a shifted pH-optimum compared to the wild-type enzyme | Escherichia coli |
G227L | site-directed mutagenesis of subunit B, the mutant shows reduced activity and a shifted pH-optimum compared to the wild-type enzyme | Escherichia coli |
G227L/D95E | site-directed mutagenesis of subunits B and C, respectively, the mutant shows a shifted pH optimum compared to the wild-type enzyme and is inactive above pH 7.0 | Escherichia coli |
G227L/D95L | site-directed mutagenesis of subunits B and C, respectively, the mutant shows a shifted pH optimum compared to the wild-type enzyme and is inactive above pH 7.0 | Escherichia coli |
Q78L | site-directed mutagenesis of subunit D, the mutant shows reduced activity and a shifted pH-optimum compared to the wild-type enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | midpoint potentials of [3Fe-4S] cluster and heme b, kinetics and kinetic isotope effects of recombinant wild-type and mutant enzymes at different pH in both reaction directions, overview | Escherichia coli | |
0.06 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHD mutant Q78L | Escherichia coli | |
0.07 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant D95L | Escherichia coli | |
0.09 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHB mutant G227L | Escherichia coli | |
0.09 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant E101L | Escherichia coli | |
0.1 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant E101D | Escherichia coli | |
0.11 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant E101L | Escherichia coli | |
0.11 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHD mutant Q78L | Escherichia coli | |
0.124 | - |
reduced plumbagin | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95L | Escherichia coli | |
0.128 | - |
reduced plumbagin | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95E | Escherichia coli | |
0.13 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHB mutant G227L | Escherichia coli | |
0.13 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant D95E | Escherichia coli | |
0.14 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant E101D | Escherichia coli | |
0.15 | - |
ubiquinone | pH 8.0, 25°C, recombinant wild-type enzyme | Escherichia coli | |
0.16 | - |
ubiquinone | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95L | Escherichia coli | |
0.16 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant D95L | Escherichia coli | |
0.17 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant wild-type enzyme | Escherichia coli | |
0.19 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant D95E | Escherichia coli | |
0.2 | - |
ubiquinone | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95E | Escherichia coli |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | - |
Escherichia coli | 16020 | - |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Fe2+ | heme cofactor and [3Fe-4S] cluster, midpoint potentials of wild-type and mutant enzymes, overview | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
succinate + ubiquinone | Escherichia coli | - |
fumarate + ubiquinol | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes partially from strain DW35 by membrane preparation | Escherichia coli |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
additional information | aerobic growth on succinate and anaerobic growth on glycerol-fumarate | Escherichia coli | - |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
catalytic efficiencies of recombinant wild-type and mutant enzymes at different pH in both reaction directions, overview | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
fumarate + reduced plumbagin | - |
Escherichia coli | succinate + oxidized plumbagin | - |
? | |
succinate + 2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | i.e. MTT, in presence of phenazine methosulfate, i.e. PMS | Escherichia coli | ? | - |
? | |
succinate + ubiquinone | - |
Escherichia coli | fumarate + ubiquinol | - |
? | |
succinate + ubiquinone | the enzyme does not generate a proton motive force during catalysis and are electroneutral, thus, the quinone reduction reaction must consume cytoplasmic protons which are released stoichiometrically during succinate oxidation. Residues SdhBG227, SdhCD95, and SdhCE101 are located at or near the entrance of a water channel that functions as a proton wire connecting the cytoplasm to the quinone binding site in vivo, while an alternative proton pathway exists in vitro only, overview | Escherichia coli | fumarate + ubiquinol | - |
? |
Synonyms | Comment | Organism |
---|---|---|
More | SDH belongs to the highly conserved complex II family of enzymes that reduce ubiquinone | Escherichia coli |
SDH | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
7.7 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant E101L | Escherichia coli | |
8.6 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant D95L | Escherichia coli | |
9.1 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHB mutant G227L | Escherichia coli | |
9.3 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHD mutant Q78L | Escherichia coli | |
9.4 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95L | Escherichia coli | |
10.4 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant E101D | Escherichia coli | |
11.6 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHB mutant G227L | Escherichia coli | |
12.6 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant subunit SDHC mutant E101L | Escherichia coli | |
12.7 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant subunit SDHC mutant D95E | Escherichia coli | |
15.6 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant E101L | Escherichia coli | |
15.9 | - |
reduced plumbagin | pH 8.0, 25°C, recombinant wild-type enzyme | Escherichia coli | |
15.9 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant subunit SDHB mutant G227L | Escherichia coli | |
16.2 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant D95L | Escherichia coli | |
17.4 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant subunit SDHC mutant D95L | Escherichia coli | |
17.4 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHD mutant Q78L | Escherichia coli | |
18 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant subunit SDHC mutant E101D | Escherichia coli | |
18.6 | - |
ubiquinone | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95E | Escherichia coli | |
19.9 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95E | Escherichia coli | |
20.8 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant subunit SDHD mutant Q78L | Escherichia coli | |
20.9 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant wild-type enzyme | Escherichia coli | |
20.9 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant E101D | Escherichia coli | |
21.5 | - |
reduced plumbagin | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95L | Escherichia coli | |
22.3 | - |
ubiquinone | pH 8.0, 25°C, recombinant subunit SDHC mutant D95E | Escherichia coli | |
23 | - |
reduced plumbagin | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95E | Escherichia coli | |
24.8 | - |
ubiquinone | pH 7.0, 25°C, recombinant subunit SDHB/C mutant G227L/D95L | Escherichia coli | |
29.6 | - |
2-(4,5-dimethyl-2-thiazolyl)-3,5-diphenyl-2H-tetrazolium bromide | pH 8.0, 25°C, recombinant subunit SDHC mutant D95E | Escherichia coli | |
37.9 | - |
ubiquinone | pH 8.0, 25°C, recombinant wild-type enzyme | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
- |
Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
flavin | quantitative determination of content in wild-type and mutant enzymes | Escherichia coli | |
heme | a single heme b moiety is incorporated into the membrane anchor and only the QP-site is functional | Escherichia coli | |
Plumbagin | a quinone analogue | Escherichia coli | |
quinone | with a periplasmically oriented quinone binding site of the enzyme | Escherichia coli |