Cloned (Comment) | Organism |
---|---|
expression of HCA II in Escherichia coli strain BL21 | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant HCA II, crystals are prepared at pH 6.0, pH 8.5, and pH 11.0, hanging drop vapor diffusion method, crystal preparation from apo-Co(II)-HCA II by mixing 0.005 ml of protein solution with 0.005 ml of precipitant solution, containing 1.4 M sodium citrate, and 100 mM Tris-HCl, pH 9.0, at room temperature, and soaking of the crystals in 100 mM CoCl2, 1.4 M sodium citrate, 50 mM Tris-Cl, 50 mM 3-(cyclohexylamino)propanesulfonic acid with pH values of 6.0, 8.5 and 11.0, respectively, 2-3days, X-ray diffraction structure determination and analysis at 1.5-1.6 A resolution | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | no inhibition of HCA II by citrate and no conformational changes of amino-acid side chains as a result of the use of citrate at pH 6.0-11.0 | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Co2+ | removal of Zn2+ and preparation of Co(II)-substituted carbonic anhydrase II, Co(II)-HCA II. Change in metal coordination from tetrahedral at pH 11.0 to a coordination consistent with a mixed population of both tetrahedral and penta-coordinate at pH 8.5 to an octahedral geometry characteristic of the oxidized enzyme Co(III)-HCA II at pH 6.0 | Homo sapiens | |
Zn2+ | required | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
H2CO3 | Homo sapiens | - |
CO2 + H2O | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P00918 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant HCA II from Escherichia coli strain BL21 | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
H2CO3 | - |
Homo sapiens | CO2 + H2O | - |
r |
Synonyms | Comment | Organism |
---|---|---|
carbonic anhydrase II | - |
Homo sapiens |
HCA II | - |
Homo sapiens |
pH Minimum | pH Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
change in metal coordination from tetrahedral at pH 11.0 to a coordination consistent with a mixed population of both tetrahedral and penta-coordinate at pH 8.5 to an octahedral geometry characteristic of the oxidized enzyme Co(III)-HCA II at pH 6.0 | Homo sapiens |
6 | 11 | - |
Homo sapiens |