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Literature summary for 7.5.2.2 extracted from

  • Benabdelhak, H.; Schmitt, L.; Horn, C.; Jumel, K.; Blight, M.A.; Holland, I.B.
    Positive co-operative activity and dimerization of the isolated ABC ATPase domain of HlyB from Escherichia coli (2005), Biochem. J., 386, 489-495.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 7.5.2.2

Inhibitors

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 [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

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 [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

Organism UniProt Comment Textmining
Escherichia coli
-
 strain DH5alpha
-
Escherichia coli DH5-alpha
-
 strain DH5alpha
-

Purification (Commentary)

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

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 and Products (Substrate)

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

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

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 [1/s]

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

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 in the presence of 3 mM MgCl2 and 10 mM KCl Escherichia coli

pH Range

pH Minimum pH Maximum Comment Organism
7 8.5
-
 Escherichia coli

pH Stability

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 IC50 Value Maximum Comment Organism Inhibitor Structure
0.016
-
 0.005-0.05 mM, pH 8.0, IC50: 0.016 mM Escherichia coli orthovanadate