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Literature summary extracted from

  • Siarheyeva, A.; Sharom, F.J.
    The ABC transporter MsbA interacts with lipid A and amphipathic drugs at different sites (2009), Biochem. J., 419, 317-328.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
7.5.2.6 expression of His6-tagged wild-type and mutant enzyme in Escherichia coli Salmonella enterica subsp. enterica serovar Typhimurium

Protein Variants

EC Number Protein Variants Comment Organism
7.5.2.6 C315S site-idrected mutagenesis, structure analysis in comparison to the wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium
7.5.2.6 C88S site-directed mutagenesis, structure analysis in comparison to the wild-type enzyme Salmonella enterica subsp. enterica serovar Typhimurium

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
7.5.2.6 additional information
-
additional information ligand binding kinetics Salmonella enterica subsp. enterica serovar Typhimurium
7.5.2.6 0.41
-
ATP pH 7.5, 37°C, unlabeled recombinant MsbA Salmonella enterica subsp. enterica serovar Typhimurium
7.5.2.6 0.52
-
ATP pH 7.5, 37°C, MIANS-labeled recombinant MsbA Salmonella enterica subsp. enterica serovar Typhimurium

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
7.5.2.6 membrane
-
Salmonella enterica subsp. enterica serovar Typhimurium 16020
-

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
7.5.2.6 65000
-
x * 65000, about, purified recombinant enzyme, mass spectrometry Salmonella enterica subsp. enterica serovar Typhimurium

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 Salmonella enterica subsp. enterica serovar Typhimurium MsbA contains two substrate-binding sites that communicate with both the nucleotide-binding domain and with each other, one is a high affinity binding site for the physiological substrate, lipid A, and the other site interacts with drugs with comparable affinity, overview ADP + phosphate + Lipid A/out
-
?
7.5.2.6 ATP + H2O + lipopolysaccharide/in Salmonella enterica subsp. enterica serovar Typhimurium MsbA is an essential ABC transporter involved in lipid A transport across the cytoplasmic membrane ADP + phosphate + lipopolysaccharide/out
-
?

Organism

EC Number Organism UniProt Comment Textmining
7.5.2.6 Salmonella enterica subsp. enterica serovar Typhimurium
-
gene msbA
-

Purification (Commentary)

EC Number Purification (Comment) Organism
7.5.2.6 recombinant His6-tagged wild-type and mutant enzyme by nickel affinity chromatography Salmonella enterica subsp. enterica serovar Typhimurium

Renatured (Commentary)

EC Number Renatured (Comment) Organism
7.5.2.6 purified recombinant His6-tagged MsbA is reconstituted into proteoliposomes, that contain phosphocholine, using detergent destabilization Salmonella enterica subsp. enterica serovar Typhimurium

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
-
protein quantification by labeling of the purified native protein with MIANS, i.e. 2-(4'-maleimidylanilino)naphthalene-6-sulfonic acid, on Cys315 Salmonella enterica subsp. enterica serovar Typhimurium
7.5.2.6 0.4
-
ATPase activity, purified recombinant enzyme, labeled or unlabeled with 2-(4'-maleimidylanilino)naphthalene-6-sulfonic acid Salmonella enterica subsp. enterica serovar Typhimurium

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
7.5.2.6 ATP + H2O + Lipid A/in MsbA contains two substrate-binding sites that communicate with both the nucleotide-binding domain and with each other, one is a high affinity binding site for the physiological substrate, lipid A, and the other site interacts with drugs with comparable affinity, overview Salmonella enterica subsp. enterica serovar Typhimurium ADP + phosphate + Lipid A/out
-
?
7.5.2.6 ATP + H2O + lipopolysaccharide/in
-
Salmonella enterica subsp. enterica serovar Typhimurium ADP + phosphate + lipopolysaccharide/out
-
?
7.5.2.6 ATP + H2O + lipopolysaccharide/in MsbA is an essential ABC transporter involved in lipid A transport across the cytoplasmic membrane Salmonella enterica subsp. enterica serovar Typhimurium ADP + phosphate + lipopolysaccharide/out
-
?
7.5.2.6 additional information binding of of amphipathic drugs, e.g. of daunorubicin, quercetin, verapamil, or propafenone GP12, alters the protein conformation Salmonella enterica subsp. enterica serovar Typhimurium ?
-
?

Subunits

EC Number Subunits Comment Organism
7.5.2.6 ? x * 65000, about, purified recombinant enzyme, mass spectrometry Salmonella enterica subsp. enterica serovar Typhimurium
7.5.2.6 More secondary structure analysis, binding of of amphipathic drugs, e.g. of daunorubicin, alters the protein conformation, topical enzyme model, overview Salmonella enterica subsp. enterica serovar Typhimurium

Synonyms

EC Number Synonyms Comment Organism
7.5.2.6 MsbA
-
Salmonella enterica subsp. enterica serovar Typhimurium

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
7.5.2.6 additional information
-
ligand binding assays at 23°C Salmonella enterica subsp. enterica serovar Typhimurium
7.5.2.6 37
-
ATPase assay at Salmonella enterica subsp. enterica serovar Typhimurium

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5.2.6 7.5
-
ATPase assay at Salmonella enterica subsp. enterica serovar Typhimurium

Cofactor

EC Number Cofactor Comment Organism Structure
7.5.2.6 ATP
-
Salmonella enterica subsp. enterica serovar Typhimurium