EC Number | Cloned (Comment) | Organism |
---|---|---|
4.2.1.1 | recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 (DE3)pLysS | Chlamydomonas reinhardtii |
EC Number | Crystallization (Comment) | Organism |
---|---|---|
4.2.1.1 | purified recombinant detagged enzyme CrCAH3 with inhibitors phosphate and acetazolamide, hanging drop vapor diffusion method, mixing of 0.001 ml of 3.6 mg/l protein in mM Tris-HCl, pH 8.0, and 150 mM NaCl, and 1 mM ligand, with 0.001 ml of reservoir solution containing 2.5 M NH4H2PO4 and 0.1 M Tris-HCl, pH 8.0, to a final pH of pH 4.1, and equilibration over 1 ml reservoir solution, at 18°C, method optimization, X-ray diffraction structure determination and analysis at 2.6-2.7 A resolution | Chlamydomonas reinhardtii |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
4.2.1.1 | additional information | generation of a CrCIA3 deletion mutant. Point mutations A399C, C499G, C525G, A570G, G732C, G888C, and C903G, none of these mutations introduces changes in the CrCAH3 amino acid sequence | Chlamydomonas reinhardtii |
EC Number | Inhibitors | Comment | Organism | Structure |
---|---|---|---|---|
4.2.1.1 | acetazolamide | binding structure analysis, overview. Each unit cell is comprised of four dimers of CrCAH3 | Chlamydomonas reinhardtii | |
4.2.1.1 | DTT | 50% inhibition at 0.5 mM, complete inhibition at above 2.5 mM | Chlamydomonas reinhardtii | |
4.2.1.1 | phosphate | binding structure analysis, overview. Phosphate binds to the catalytic zinc cofactor, each unit cell is comprised of four dimers of CrCAH3 | Chlamydomonas reinhardtii |
EC Number | Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|---|
4.2.1.1 | additional information | enzyme CrCAH3 is associated with photosystem II, PSII | Chlamydomonas reinhardtii | - |
- |
4.2.1.1 | thylakoid lumen | alpha-type CA (CrCAH3) locates in the thylakoid lumen | Chlamydomonas reinhardtii | 31977 | - |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
4.2.1.1 | Zn2+ | required, metalloenzyme | Chlamydomonas reinhardtii |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.1.1 | H2CO3 | Chlamydomonas reinhardtii | - |
CO2 + H2O | - |
r |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
4.2.1.1 | Chlamydomonas reinhardtii | A8J4Z8 | cell wall-deficient Chlamydomonas reinhardtii mutant92 (cw92), referred to as wild-type | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.2.1.1 | recombinant is-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 (DE3)pLysS by nickel affinity chromatography, gel filtration, and tag cleavage by TEV protease, followed by dialysis, another step of nickel affinity chromatography, anion exchange chromatography, gel filtration, and ultrafiltration | Chlamydomonas reinhardtii |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
4.2.1.1 | H2CO3 | - |
Chlamydomonas reinhardtii | CO2 + H2O | - |
r |
EC Number | Subunits | Comment | Organism |
---|---|---|---|
4.2.1.1 | dimer | enzyme CrCAH3 is a dimer at pH 4.1 that is stabilized by swapping of the N-terminal arms, a feature not previously observed in alpha-type carbonic anhydrases | Chlamydomonas reinhardtii |
4.2.1.1 | More | each unit cell is comprised of four dimers of CrCAH3 | Chlamydomonas reinhardtii |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
4.2.1.1 | alpha-type CA | - |
Chlamydomonas reinhardtii |
4.2.1.1 | CAH3 | - |
Chlamydomonas reinhardtii |
4.2.1.1 | CrCAH3 | - |
Chlamydomonas reinhardtii |
4.2.1.1 | luminal carbonic anhydrase | - |
Chlamydomonas reinhardtii |
4.2.1.1 | photosystem II-associated carbonic anhydrase | - |
Chlamydomonas reinhardtii |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
4.2.1.1 | 20 | - |
assay at | Chlamydomonas reinhardtii |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
4.2.1.1 | 6 | 6.5 | - |
Chlamydomonas reinhardtii |
EC Number | General Information | Comment | Organism |
---|---|---|---|
4.2.1.1 | malfunction | a CrCIA3 deletion mutant shows 50% reduction of the O2 evolution rate | Chlamydomonas reinhardtii |
4.2.1.1 | additional information | a disulfide bond between Cys90 and Cys258 is essential for CrCAH3 activity | Chlamydomonas reinhardtii |
4.2.1.1 | physiological function | in the green alga Chlamydomonas reinhardtii, a luminal carbonic anhydrase, CrCAH3 improves proton removal from phorosystem PSII, possibly by rapid reformation of HCO3- from CO2. It plays a direct role of CrCAH3 in the turnover efficiency of PSII, and a stimulating effect of CrCAH3 and CO2/HCO3- on PSII activity. Possible redox regulation of the enzyme, overview. Redox regulation in the chloroplast thylakoid lumen is a common way to regulate lumenal and photosynthetic proteins | Chlamydomonas reinhardtii |