Inhibitors | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | 3% activity at pH 8.0 and 10 mM KCl | Escherichia coli | |
Co2+ | 47% activity at pH 8.0 and 10 mM KCl | Escherichia coli | |
additional information | the activity is reversibly inhibited by high salt concentrations in the physiologically range accompanied by proportional decreased binding of 8-azido-ATP (3-5% residual activity at 200-300 mM salt) | Escherichia coli | |
orthovanadate | 0.005-0.05 mM, pH 8.0, IC50: 0.016 mM | Escherichia coli | |
Zn2+ | 1% activity at pH 8.0 and 10 mM KCl | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.65 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 10 mM NaCl | Escherichia coli | |
1.09 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 50 mM NaCl | Escherichia coli | |
1.5 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 100 mM NaCl | Escherichia coli | |
2.33 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 200 mM NaCl | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | 100% activity at pH 8.0 and with 10 mM KCl | Escherichia coli | |
Mn2+ | 125% activity at pH 8.0 and with 10 mM KCl | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
27500 | - |
HlyB NBD in the presence of 100 mM KCl and ATP, ultracentrifugation | Escherichia coli |
27700 | - |
HlyB NBD in the presence of 10 mM KCl and in the absence of ATP, ultracentrifugation | Escherichia coli |
28000 | - |
His-tagged HlyB ABC domain, gel filtration | Escherichia coli |
30800 | - |
HlyB NBD in the presence of 10 mM KCl and ATP, ultracentrifugation | Escherichia coli |
35700 | - |
HlyB NBD in the absence of KCl and ATP, ultracentrifugation | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
strain DH5alpha | - |
Escherichia coli DH5-alpha | - |
strain DH5alpha | - |
Purification (Comment) | Organism |
---|---|
HlyB ABC domain, Ni2+-chelating Sepharose 16/20 fast flow column chromatography, His-tagged HlyB ABC domain, immobilized metal-affinity chromatography | Escherichia coli |
Storage Stability | Organism |
---|---|
-80°C, 50 mM phosphate buffer pH 8, 50 mM KCl and 10% glycerol, 15 mg/ml HlyB ABC domain, indefinite storage duration, no loss of activity | Escherichia coli |
-80°C, phosphate or Tris buffer pH 8, in the absence of ATP, up to 25 mg/ml His-tagged HlyB ABC domain, indefinite storage duration, no loss of activity | Escherichia coli |
25°C, 50 mM phosphate buffer pH 8, 50 mM KCl and 10% glycerol, 15 mg/ml HlyB ABC domain, 24h, 20% loss of activity | Escherichia coli |
4°C, 50 mM phosphate buffer pH 8, 50 mM KCl and 10% glycerol, 15 mg/ml HlyB ABC domain, 2 days, no loss of activity | Escherichia coli |
4°C, phosphate or Tris buffer pH 8, in the absence of ATP, up to 25 mg/ml His-tagged HlyB ABC domain, at least 2 days, no loss of activity | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
8-azido-ATP + H2O + ? | - |
Escherichia coli | 8-azido-ADP + phosphate + ? | - |
? | |
8-azido-ATP + H2O + ? | - |
Escherichia coli DH5-alpha | 8-azido-ADP + phosphate + ? | - |
? | |
ATP + H2O + ? | - |
Escherichia coli | ADP + phosphate + ? | - |
? | |
ATP + H2O + ? | - |
Escherichia coli DH5-alpha | ADP + phosphate + ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 28000, gel filtration, small amounts of dimers (apparently when stabilized by 8-azido-ATP) are detected even in the presence of salt | Escherichia coli |
monomer | 1 * 28000, gel filtration and ultracentrifugation, over a wide range of protein or NaCl or KCl concentrations the enzyme is only detected as monomer | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
ABC ATPase domain of HlyB transporter | - |
Escherichia coli |
ATP-binding cassette ATPase domain of the haemolysin B transporter | - |
Escherichia coli |
HlyB NBD | - |
Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.11 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 200 mM NaCl | Escherichia coli | |
0.2 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 100 mM NaCl | Escherichia coli | |
0.31 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 50 mM NaCl | Escherichia coli | |
0.33 | - |
ATP | KM-value with 50% of the enzyme being substrate-saturated, in the presence of 10 mM NaCl | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
in the presence of 3 mM MgCl2 and 10 mM KCl | Escherichia coli |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
7 | 8.5 | - |
Escherichia coli |
pH Stability | pH Stability Maximum | Comment | Organism |
---|---|---|---|
7 | - |
75% activity in the presence of 3 mM MgCl2 and 10 mM KCl | Escherichia coli |
7.5 | - |
90% activity in the presence of 3 mM MgCl2 and 10 mM KCl | Escherichia coli |
8 | - |
100% activity in the presence of 3 mM MgCl2 and 10 mM KCl | Escherichia coli |
8.5 | - |
85% activity in the presence of 3 mM MgCl2 and 10 mM KCl | Escherichia coli |
IC50 Value | IC50 Value Maximum | Comment | Organism | Inhibitor | Structure |
---|---|---|---|---|---|
0.016 | - |
0.005-0.05 mM, pH 8.0, IC50: 0.016 mM | Escherichia coli | orthovanadate |