CAH3 | Carbonic anhydrase
AS05 073 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Chlamydomonas reinhardtii
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Recombinant carbonic anhydrase of Chlamydomonas reinhardtii A8J4Z8 after cleavage of a fusion protein
33.4 | 29 kDa
Chlamydomonas reinhardtii (only)
Arabidopsis thaliana, Oryza sativa
(1) Chlamydomonas reinhardtii thylakoid preparation, (2) overexoressed carbonic anhydrase, primary antibody used in 1: 2000 dilution. Secondary antibodies anti-rabbit IgG. used at 1: 10 000 with chemiluminescent detection.
20 µg/ml of chlorophyll from Chlamydomonas reinhardtii Cah3-less mutant (1), and 15 µg/ml of chlorophyll from Chlamydomonas reinhardtii wild-type (2),extracted with 4x Laemmli were separated on 12 % SDS-PAGE and blotted 2h to nitrocellulose filter using tank transfer. Blots were blocked with 5% milk powder for for 2h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 fover night at 8°C with agitation. The antibody solution was decanted and the blot was rinsed briefly, then washed once for 15 min and 3 times for 20 min each in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in for 1h at RT with agitation. The blot was washed as above and developed for 1 min with chemiluminescent detection reagent, according to the manufacturer's instructions. Exposure time was 10min. MW standards used, Page ruler, pre-stained from Thermofisher.
Courtesy of Dr. Göran Samuelsson, Umeå Plant Science Centre, Sweden
CAH3 - carbonic anhydrase is a zinc-containing metalloenzyme that catalyzes the reversible interconversion of CO2 and HCO3?. It plays an important role in many physiological functions that involve decarboxylation or carboxylation reactions, including both photosynthesis and respiration.
Terentyev (2020: The Main Structural and Functional Characteristics of Photosystem-II-Enriched Membranes Isolated From Wild Type and cia3 Mutant Chlamydomonas reinhardtii. Life (Basel). 2020 May 14;10(5):E63. doi: 10.3390/life10050063.
Terentyev et al. (2019). Carbonic anhydrase CAH3 supports the activity of photosystem II under increased pH. Biochim Biophys Acta Bioenerg. 2019 Jul 1;1860(7):582-590. doi: 10.1016/j.bbabio.2019.06.003.
Correa-Galvis et al. (2016). Photosystem II Subunit PsbS Is Involved in the Induction of LHCSR Protein-dependent Energy Dissipation in Chlamydomonas reinhardtii. J Biol Chem. 2016 Aug 12;291(33):17478-87. doi: 10.1074/jbc.M116.737312.
Mitchell et al. (2014). Dynamics of carbon concentrating mechanism induction and protein re-localisation during the dark to light transition in synchronised Chlamydomonas. Plant Physiol. 2014 Aug 8. pii: pp.114.246918.
Tirumani et al. (2014). Regulation of CCM genes in Chlamydomonas reinhardtii during conditions of light-dark cycles in synchronous cultures. Plant Mol Biol. 2014 Mar 4.
Shutova et al. (2008). The photosystem II-associated Cah3 in Chlamydomonas enhances the O2evolution rate by proton removal. EMBO J. 27:782-791.
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