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Literature summary for 7.5.2.6 extracted from

  • Eckford, P.D.; Sharom, F.J.
    The reconstituted Escherichia coli MsbA protein displays lipid flippase activity (2010), Biochem. J., 429, 195-203 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 7.5.2.6

Cloned(Commentary)

Cloned (Comment) Organism
gene msbA, overexpression of N-terminally His6-tagged enzyme in Escherichia coli Escherichia coli

Inhibitors

Inhibitors Comment Organism Structure
lipid A translocation of NBD-phosphatidylethanolamine is inhibited by the presence of the putative physiological substrate lipid A, probably due to competition for flipping of 7-nitrobenz-2-oxa-1,3-diazole-labeled phosphoethanolamine (NBD-PE) Escherichia coli

Localization

Localization Comment Organism GeneOntology No. Textmining
membrane
-
 Escherichia coli 16020
-

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + H2O + lipid A-core oligosaccharide[side 1] Escherichia coli
-
 ADP + phosphate + lipid A-core oligosaccharide[side 2]
-
 ?

Organism

Organism UniProt Comment Textmining
Escherichia coli P60752
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant N-terminally His6-tagged enzyme from isolated Escherichia coli inside-out membrane vesicles by nickel affinity chromatography Escherichia coli

Renatured (Commentary)

Renatured (Comment) Organism
reconstitution of purified MsbA into proteoliposomes, method, overview Escherichia coli

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + H2O + lipid A-core oligosaccharide[side 1]
-
 Escherichia coli ADP + phosphate + lipid A-core oligosaccharide[side 2]
-
 ?
ATP + H2O + phosphoethanolamine[side 1] fluorescently NBD (7-nitrobenz-2-oxa-1,3-diazole)-labeled Escherichia coli ADP + phosphate + phosphoethanolamine[side 2]
-
 ?
additional information purified MsbA from Escherichia coli displays high ATPase activity, and binds to lipids and lipid-like molecules, including lipid A, with affinity in the low micromolar range. Purified MsbA is reconstituted into proteoliposomes of Escherichia coli lipid and shows ability to translocate 7-nitrobenz-2-oxa-1,3-diazole (NBD)-labeled lipid derivatives. 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 an acyl chain-labeled phosphatidylethanolamine derivative. Lipid flippase activity requires ATP hydrolysis, and is dependent on the concentration of ATP and NBD-lipid. MsbA can accommodate lipids with bulky carbohydrate headgroups Escherichia coli ?
-
 ?

Subunits

Subunits Comment Organism
homodimer
-
 Escherichia coli

Synonyms

Synonyms Comment Organism
lipid flippase
-
 Escherichia coli
MsbA
-
 Escherichia coli

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
 assay at Escherichia coli

General Information

General Information Comment Organism
evolution the MsbA protein is an essential ABC (ATP-binding-cassette) superfamily member in Gram-negative bacteria Escherichia coli
additional information reconstitution of purified MsbA into proteoliposomes, method, overview Escherichia coli
physiological function the homodimeric ATP-dependent lipid translocase or flippase transports lipid A from the inner to the outer leaflet of the cytoplasmic membrane Escherichia coli