Literature summary extracted from
Cooper, R.S.; Altenberg, G.A.
Association/dissociation of the nucleotide-binding domains of the ATP-binding cassette protein MsbA measured during continuous hydrolysis (2013), J. Biol. Chem., 288, 20785-20796 .
Cloned(Commentary)
EC Number |
Cloned (Comment) |
Organism |
---|
7.5.2.6 |
gene msbA, recombinant expression of His10-tagged wild-type and mutant enzymes in Escherichia coli strain BL21-Codon plus(DE3)-RIL |
Salmonella enterica subsp. enterica serovar Typhimurium |
Protein Variants
EC Number |
Protein Variants |
Comment |
Organism |
---|
7.5.2.6 |
C315S |
site-directed mutagenesis of the active site Cys |
Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 |
C88A/C315A/E506Q/T561C |
site-directed mutagenesis, replacement of the catalytic carboxylate E506, the catalytically deficient mutant E506Q/T561C has a very low activity (1% of the T561C activity) |
Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 |
C88A/C315A/T561C |
site-directed mutagenesis, inactive cysteine mutant, the ATPase activity of T561C is highly dependent on temperature, addition of Mg2x01 decreased the LRET signal intensity of the ATP-bound T561C, and subsequent addition of Vi increased the signal back toward the ATP-bound state |
Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 |
C88S |
site-directed mutagenesis of the active site Cys |
Salmonella enterica subsp. enterica serovar Typhimurium |
7.5.2.6 |
additional information |
the mutant enzymes are T561C and E506Q/T561C are labeled with thiol-reactive LRET probes, time course of nucleotide-binding domains (NBDs) association/dissociation monitored by LRET, kinetics of the conformational changes |
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 |
kinetics of the conformational change from ATP-bound T561C to the dynamic equilibrium during continuous ATP hydrolysis (in MgATP) by stopped flow measurements, kinetics of the conformational changes of mutant enzymes |
Salmonella enterica subsp. enterica serovar Typhimurium |
|
Metals/Ions
EC Number |
Metals/Ions |
Comment |
Organism |
Structure |
---|
7.5.2.6 |
Mg2+ |
required, residue Glu506, located at the end of motif B, is the catalytic carboxylate involved in Mg2+ coordination |
Salmonella enterica subsp. enterica serovar Typhimurium |
|
Molecular Weight [Da]
EC Number |
Molecular Weight [Da] |
Molecular Weight Maximum [Da] |
Comment |
Organism |
---|
7.5.2.6 |
134000 |
- |
gel filtration |
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-core oligosaccharide[side 1] |
Salmonella enterica subsp. enterica serovar Typhimurium |
- |
ADP + phosphate + lipid A-core oligosaccharide[side 2] |
- |
? |
|
Organism
EC Number |
Organism |
UniProt |
Comment |
Textmining |
---|
7.5.2.6 |
Salmonella enterica subsp. enterica serovar Typhimurium |
P63359 |
- |
- |
Purification (Commentary)
EC Number |
Purification (Comment) |
Organism |
---|
7.5.2.6 |
recombinant His10-tagged wild-type and mutant enzymes from Escherichia coli strain BL21-Codon plus(DE3)-RIL by solubilization from membranes with 2% n-dodecyl-beta-D-maltopyranoside and 0.4% cholate, cobalt affinity chromatography, and gel filtration in presence of detergents |
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-core oligosaccharide[side 1] |
- |
Salmonella enterica subsp. enterica serovar Typhimurium |
ADP + phosphate + lipid A-core oligosaccharide[side 2] |
- |
? |
|
Subunits
EC Number |
Subunits |
Comment |
Organism |
---|
7.5.2.6 |
homodimer |
- |
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 |
37 |
- |
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 |
- |
assay at |
Salmonella enterica subsp. enterica serovar Typhimurium |
Cofactor
EC Number |
Cofactor |
Comment |
Organism |
Structure |
---|
7.5.2.6 |
ATP |
induces conformational changes of the enzyme during catalysis, kinetics of the conformational changes of mutant enzymes, overview |
Salmonella enterica subsp. enterica serovar Typhimurium |
|
General Information
EC Number |
General Information |
Comment |
Organism |
---|
7.5.2.6 |
additional information |
structure of the AMPPNP-bound MsbA, structure-activity model, overview |
Salmonella enterica subsp. enterica serovar Typhimurium |