EC Number | Cloned (Comment) | Organism |
---|---|---|
7.5.2.6 | gene msbA | Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 | gene msbA, recombinant overexpresssion of FLAG-tagged enzyme in Escherichia coli strain Rosetta 2 (DE3) | Escherichia coli |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
7.5.2.6 | purified enzyme MsbA free and in complex with inhibitors (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid or G902, sitting drop vapour diffusion method, mixing of 10 mg/ml protein in 50 mM Tris, pH 8.0, 500 mM NaCl, and 0.6% FA-3 detergent, with mother liquor containing 200 mM NaCl, 19% PEG 550 MME, 4% PEG 400, and 100 mM HEPES, pH 7.0, in a 1:1 v/v ratio, 19°C, 1 week, X-ray diffraction structure determination and analysis at 2.9 A resolution | Escherichia coli |
7.5.2.6 | purified enzyme MsbA in complex with inhibitor G907, analysis of structure PDB ID 3B60 | Salmonella enterica subsp. enterica serovar Typhimurium |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
7.5.2.6 | additional information | overexpression of msbA suppresses mutations in the htrB lipid A acyltransferase, as MsbA can transport the tetra-acylated LPS produced in htrB mutants, albeit inefficiently | Escherichia coli |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
7.5.2.6 | (2E)-3-[1-cyclopropyl-7-[(1S)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]naphthalen-2-yl]prop-2-enoic acid | - |
Escherichia coli | |
7.5.2.6 | (2E)-3-[1-cyclopropyl-7-[(1S)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]naphthalen-2-yl]prop-2-enoic acid | - |
Salmonella enterica subsp. enterica serovar Typhimurium | |
7.5.2.6 | (2E)-3-[6-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]quinolin-3-yl]prop-2-enoic acid | - |
Escherichia coli | |
7.5.2.6 | (2E)-3-[6-[(1R)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]quinolin-3-yl]prop-2-enoic acid | - |
Salmonella enterica subsp. enterica serovar Typhimurium | |
7.5.2.6 | (2E)-3-[6-[(1S)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]quinolin-3-yl]prop-2-enoic acid | a quinoline compound with potent activity on purified Escherichia coli MsbA | Escherichia coli | |
7.5.2.6 | (2E)-3-[6-[(1S)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]quinolin-3-yl]prop-2-enoic acid | - |
Salmonella enterica subsp. enterica serovar Typhimurium | |
7.5.2.6 | (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid | selective small-molecule antagonist with bactericidal activity and a dual-mode inhibitory mechanism. (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid traps MsbA in an inward-facing, lipopolysaccharide-bound conformation by wedging into an architecturally conserved transmembrane pocket. (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid gains access to MsbA through the bulk membrane. The 2-chloro-6 cyclopropylphenyl substituent of (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid (A-ring) exhibits strong electron density and makes van der Waals interactions with side chains from transmembrane domains TM4 (L171, A175 and V178), TM5 (A259 and L263) and TM6 (M291 and L294). The quinoline core of (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid (B-ring) is orthogonal to the plane of the phenyl substituent, where it is partially enclosed by residues from transmembrane domains TM4 (V178, S179 and I182), TM5 (A259) and TM6 (M295 and L298). A259 and M295 side chains contact the 4-cyclopropyl substitution of the quinoline core and delineate the inhibitor binding-pocket from the inner vestibule of MsbA. (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid stabilizes a catalytically incompetent state of the transporter. The inhibitor may have broad relevance across the ABC transporter superfamily | Escherichia coli | |
7.5.2.6 | (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid | i.e. G907, selective small-molecule antagonist with bactericidal activity and a dual-mode inhibitory mechanism. G907-MsbA binding structure, overview. In the outward-facing StMsbA structure, the G907 pocket is markedly deformed and expected to perturb inhibitor binding. This substantial perturbation of the binding site during the transport cycle defines G907 as an inward-facing state-dependent inhibitor. The inhibitor may have broad relevance across the ABC transporter superfamily | Salmonella enterica subsp. enterica serovar Typhimurium | |
7.5.2.6 | additional information | analysis of ABC transporter inhibition mechanism, small molecule inhibitor library screening, overview. (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid traps MsbA in an inward-facing, lipopolysaccharide-bound conformation by wedging into an architecturally conserved transmembrane pocket. A second allosteric mechanism of antagonism occurs through structural and functional uncoupling of the nucleotide-binding domains | Escherichia coli | |
7.5.2.6 | additional information | analysis of ABC transporter inhibition mechanism, small molecule inhibitor library screening, overview | Salmonella enterica subsp. enterica serovar Typhimurium |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
7.5.2.6 | periplasm | - |
Salmonella enterica subsp. enterica serovar Typhimurium | - |
- |
7.5.2.6 | periplasm | - |
Escherichia coli | - |
- |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
7.5.2.6 | Mg2+ | required | Salmonella enterica subsp. enterica serovar Typhimurium | |
7.5.2.6 | Mg2+ | required | Escherichia coli |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.5.2.6 | ATP + H2O + lipid A-core oligosaccharide[side 1] | Salmonella enterica subsp. enterica serovar Typhimurium | - |
ADP + phosphate + lipid A-core oligosaccharide[side 2] | - |
? | |
7.5.2.6 | ATP + H2O + lipid A-core oligosaccharide[side 1] | Escherichia coli | - |
ADP + phosphate + lipid A-core oligosaccharide[side 2] | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
7.5.2.6 | Escherichia coli | P60752 | - |
- |
7.5.2.6 | Salmonella enterica subsp. enterica serovar Typhimurium | P63359 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
7.5.2.6 | recombinant FLAG-tagged enzyme from Escherichia coli strain Rosetta 2 (DE3) by affinity chromatography, the Flag tag is removed using TEV protease, followed by gel filtration and ultrafiltration | Escherichia coli |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
7.5.2.6 | ATP + H2O + lipid A-core oligosaccharide[side 1] | - |
Salmonella enterica subsp. enterica serovar Typhimurium | ADP + phosphate + lipid A-core oligosaccharide[side 2] | - |
? | |
7.5.2.6 | ATP + H2O + lipid A-core oligosaccharide[side 1] | - |
Escherichia coli | ADP + phosphate + lipid A-core oligosaccharide[side 2] | - |
? | |
7.5.2.6 | additional information | MsbA cannot efficiently transport a substrate lacking phosphorylation at the 4'-position of lipid A | Salmonella enterica subsp. enterica serovar Typhimurium | ? | - |
? | |
7.5.2.6 | additional information | MsbA cannot efficiently transport a substrate lacking phosphorylation at the 4'-position of lipid A | Escherichia coli | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
7.5.2.6 | EcMsbA | - |
Escherichia coli |
7.5.2.6 | lipid A export ATP-binding/permease protein MsbA | UniProt | Escherichia coli |
7.5.2.6 | MsbA | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 | MsbA | - |
Escherichia coli |
7.5.2.6 | STM0984 | locus name | Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 | StMsbA | - |
Salmonella enterica subsp. enterica serovar Typhimurium |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
7.5.2.6 | 37 | - |
assay at | Escherichia coli |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
7.5.2.6 | 7.5 | - |
assay at | Escherichia coli |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
7.5.2.6 | ATP | - |
Salmonella enterica subsp. enterica serovar Typhimurium | |
7.5.2.6 | ATP | - |
Escherichia coli |
EC Number | IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|---|
7.5.2.6 | 0.000005 | - |
pH 7.5, 37°C, recombinant detagged enzyme | Escherichia coli | (2E)-3-[1-cyclopropyl-7-[(1S)-1-(2,6-dichloro-3-fluorophenyl)ethoxy]naphthalen-2-yl]prop-2-enoic acid | |
7.5.2.6 | 0.000018 | - |
pH 7.5, 37°C, recombinant detagged enzyme | Escherichia coli | (2E)-3-[6-[1-(2-chloro-6-cyclopropylphenyl)ethoxy]-4-cyclopropylquinolin-3-yl]prop-2-enoic acid |
EC Number | General Information | Comment | Organism |
---|---|---|---|
7.5.2.6 | metabolism | the enzyme is involved in the lipid A biosynthesis. MsbA cannot efficiently transport a substrate lacking phosphorylation at the 4'-position of lipid A. This last essential step of lipid A biosynthesis is catalysed by LpxK, the two proteins are concatenated in some bacteria, suggesting tight coupling of their cellular activities | Escherichia coli |
7.5.2.6 | additional information | inward facing structure of EcMsbA in complex with LPS, structure comparisons and molecular basis of active lipid transport, overview | Escherichia coli |
7.5.2.6 | additional information | outward facing structure of Salmonella typhimurium MsbA (StMsbA) in complex with LPS, structure comparisons and molecular basis of active lipid transport, overview | Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 | physiological function | the movement of core-lipopolysaccharide across the inner membrane of Gram-negative bacteria is catalysed by an essential ATP-binding cassette transporter, MsbA, structure of LPS in complex with EcMsbA, overview | Escherichia coli |
7.5.2.6 | physiological function | the movement of core-lipopolysaccharide across the inner membrane of Gram-negative bacteria is catalysed by an essential ATP-binding cassette transporter, MsbA, structure of LPS in complex with StMsbA, overview | Salmonella enterica subsp. enterica serovar Typhimurium |