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Literature summary extracted from

  • Benlloch, R.; Shevela, D.; Hainzl, T.; Grundstroem, C.; Shutova, T.; Messinger, J.; Samuelsson, G.; Sauer-Eriksson, A.E.
    Crystal structure and functional characterization of photosystem II-associated carbonic anhydrase CAH3 in Chlamydomonas reinhardtii (2015), Plant Physiol., 167, 950-962 .
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

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

Crystallization (Commentary)

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

Protein Variants

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

Inhibitors

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

Localization

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
-

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.1.1 Zn2+ required, metalloenzyme Chlamydomonas reinhardtii

Natural Substrates/ Products (Substrates)

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

Organism

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
-

Purification (Commentary)

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

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.1 H2CO3
-
Chlamydomonas reinhardtii CO2 + H2O
-
r

Subunits

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

Synonyms

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

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.1 20
-
assay at Chlamydomonas reinhardtii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.1 6 6.5
-
Chlamydomonas reinhardtii

General Information

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