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Literature summary extracted from
- The reconstituted Escherichia coli MsbA protein displays lipid flippase activity (2010), Biochem. J., 429, 195-203.
Activating Compound
| EC Number | Activating Compound | Comment | Organism | Structure |
|---|---|---|---|---|
| 7.6.2.1 | ATP | flippase activity is essentially supported only by ATP. If the ATP concentration in the assay falls below 5 mM, there is a significant decrease in flippase activity. Maximal translocation occurs at 10 mM ATP | Escherichia coli |  |
| 7.6.2.1 | additional information | GTP, AMP and AMP-PNP yield low levels of translocation compared to ATP | Escherichia coli |
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 7.5.2.6 | expressed as a His-tagged fusion protein in Escherichia coli | Escherichia coli |
Inhibitors
| EC Number | Inhibitors | Comment | Organism | Structure |
|---|---|---|---|---|
| 7.6.2.1 | AlFx | trapping of MsbA with 0.8 mM AlFx results in incomplete inhibition of activity | Escherichia coli | |
| 7.6.2.1 | BeFx | trapping of MsbA with 0.8 mM BeFx results in incomplete inhibition of activity | Escherichia coli | |
| 7.6.2.1 | lipid A | lipid A inhibits translocation by approx. 30% at 0.01-0.04 mg/ml | Escherichia coli | |
| 7.6.2.1 | additional information | neither lipopolysaccharide from the Ra mutant of Escherichia coli nor deep rough chemotype lipopolysaccharide affect MsbA flippase activity significantly | Escherichia coli | |
| 7.6.2.1 | phosphate analogue Vi | the presence of 0.1-0.2 m M Vi inhibits flippase activity of MsbA by approx. 50% | Escherichia coli | |
Localization
| EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
|---|---|---|---|---|---|
| 7.6.2.1 | membrane | - |
Escherichia coli | 16020 | - |
Molecular Weight [Da]
| EC Number | Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|---|
| 7.6.2.1 | 65000 | - |
2 * 65000, SDS-PAGE | Escherichia coli |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 7.5.2.6 | ATP + H2O + Lipid A/in | Escherichia coli | - |
ADP + phosphate + Lipid A/out | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 7.5.2.6 | Escherichia coli | - |
- |
- |
| 7.6.2.1 | Escherichia coli | P60752 | - |
- |
Purification (Commentary)
| EC Number | Purification (Comment) | Organism |
|---|---|---|
| 7.6.2.1 | Ni-NTA column chromatography | Escherichia coli |
Specific Activity [micromol/min/mg]
| EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|---|
| 7.5.2.6 | additional information | - |
purified MsbA is reconstituted into proteoliposomes of Escherichia coli lipid. In this system, the protein displays maximal lipid flippase activity of 7.7 nmol of lipid translocated per mg of protein over a 20 min period for (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine (16:0, 6:0) | Escherichia coli |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-C12-sphingomyelin/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-C12-sphingomyelin/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-glucosylceramide/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-glucosylceramide/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-lactosylceramide/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-lactosylceramide/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylcholine (16:0, 6:0)/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylcholine (16:0, 6:0)/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine (16:0, 6:0)/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species. Translocation of (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine is inhibited by the presence of the putative physiological substrate lipid A | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine (16:0, 6:0)/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine (18:1)/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species. Translocation of (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine is inhibited by the presence of the putative physiological substrate lipid A | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylethanolamine (18:1)/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylglycerol (16:0, 6:0)/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylglycerol (16:0, 6:0)/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylserine (16:0, 6:0)/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylserine (16:0, 6:0)/out | - |
? | |
| 7.5.2.6 | ATP + H2O + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylserine (18:1)/in | bacterial membrane vesicles isolated from Escherichia coli overexpressing MsbA display ATP-dependent translocation of several fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labelled phospholipid species | Escherichia coli | ADP + phosphate + (7-nitrobenz-2-oxa-1,3-diazole)-phosphatidylserine (18:1)/out | - |
? | |
| 7.5.2.6 | ATP + H2O + Lipid A/in | - |
Escherichia coli | ADP + phosphate + Lipid A/out | - |
? | |
| 7.6.2.1 | ATP + H2O + Lipid A/in | - |
Escherichia coli | ADP + phosphate + Lipid A/out | - |
? | |
| 7.6.2.1 | ATP + H2O + phosphatidylethanolamine/in | - |
Escherichia coli | ADP + phosphate + phosphatidylethanolamine/out | - |
? | |
Subunits
| EC Number | Subunits | Comment | Organism |
|---|---|---|---|
| 7.6.2.1 | homodimer | 2 * 65000, SDS-PAGE | Escherichia coli |
Synonyms
| EC Number | Synonyms | Comment | Organism |
|---|---|---|---|
| 7.5.2.6 | lipid flippase | - |
Escherichia coli |
| 7.5.2.6 | MsbA | - |
Escherichia coli |
| 7.6.2.1 | lipid flippase | - |
Escherichia coli |
| 7.6.2.1 | MsbA | - |
Escherichia coli |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 7.5.2.6 | 37 | - |
assay at | Escherichia coli |
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