Cloned (Comment) | Organism |
---|---|
overexpression of HICA in Escherichia coli in minimal media supplemented with CoCl2 | Haemophilus influenzae |
Crystallization (Comment) | Organism |
---|---|
purified recombinant Co-HICA by hanging drop vapor diffusion, 10 mg/ml protein crystallized in 0.2 M sodium acetate, 0.1 M Tris-HCl, pH 8.5, 0.1 M (NH4)2SO4, and 27% PEG 4000, 22°C, several days, X-ray diffraction structure determination and analysis at 2.5 A resolution | Haemophilus influenzae |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
bicarbonate | is hypothesized to be an allosteric inhibitor of HICA | Haemophilus influenzae |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | steady-state kinetic analysis of the Co2+-substituted enzyme, overview | Haemophilus influenzae |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | preparation of a Co(II)-substituted HICA, Co-HICA. Co(II)-substituted HICA, Co-HICA, has comparable, 20% enhanced kcat and 2.3fold increased Km/kcat compared to that of the wild-type enzyme | Haemophilus influenzae | |
Zn2+ | required, can be substituted by Co2+ | Haemophilus influenzae |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
100000 | - |
about, recombinant Co-HICA, gel filtration | Haemophilus influenzae |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
H2CO3 | Haemophilus influenzae | - |
CO2 + H2O | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Haemophilus influenzae | - |
- |
- |
Purification (Comment) | Organism |
---|---|
recombinant Co-HICA from Escherichia coli by gel filtration | Haemophilus influenzae |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
H2CO3 | - |
Haemophilus influenzae | CO2 + H2O | - |
r |
Subunits | Comment | Organism |
---|---|---|
More | overall secondary and tertiary structure of Co-HICA differ from wild-type Zn-enzyme structure, active R-state and inactive T-state conformations, overview | Haemophilus influenzae |
tetramer | recombinant Co-HICA, the crystal structure shows a four-coordinate geometry for Co-HICA with pH-dependent changes in the absorption spectrum of Co-HICA, overview | Haemophilus influenzae |
Synonyms | Comment | Organism |
---|---|---|
beta-carbonic anhydrase | - |
Haemophilus influenzae |
carbonate hydrolyase | - |
Haemophilus influenzae |
HICA | - |
Haemophilus influenzae |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Haemophilus influenzae |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
and above | Haemophilus influenzae |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
6.5 | 10.5 | pH profile, pH-dependent changes in the absorption spectrum of Co-HICA including an increase in molar absorptivity and a red shift of a 580 nm peak with decreasing pH, correlate with the pH-dependence of kcat/Km, overview | Haemophilus influenzae |