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Intracellular carbonic anhydrase of Chlamydomonas reinhardtii
Umeå University, Faculty of Science and Technology, Department of Plant Physiology. Umeå University, Faculty of Science and Technology, Umeå Plant Science Centre (UPSC).ORCID iD: 0000-0002-6002-929X
Umeå University, Faculty of Science and Technology, Department of Plant Physiology.
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1995 (English)In: Plant Physiology, ISSN 0032-0889, E-ISSN 1532-2548, Vol. 109, no 2, 533-539 p.Article in journal (Refereed) Published
Abstract [en]

An intracellular carbonic anhydrase (CA; EC was purified to homogeneity from a mutant strain of Chlamydomonas reinhardtii (CW 92) lacking a cell wall. Intact cells were washed to remove periplasmic CA and were lysed and fractionated into soluble and membrane fractions by sedimentation. All of the CA activity sedimented with the membrane fraction and was dissociated by treatment with a buffer containing 200 mM KCI. Solubilized proteins were fractionated by ammonium sulfate precipitation, anionic exchange chromatography, and hydrophobic interaction chromatography. The resulting fraction had a specific activity of 1260 Wilbur-Anderson units/mg protein and was inhibited by acetazolamide (50% inhibition concentration, 12 nM). Final purification was accomplished by the specific absorption of the enzyme to a Centricon-10 microconcentrator filter. A single, 29.5-kD polypeptide was eluted from the filter with sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer, and a 1.5 M ammonium sulfate eluate contained CA activity. In comparison with human CA isoenzyme II, the N-terminal and internal amino acid sequences from the 29.5-kD polypeptide were 40% identical with the N-terminal region and 67% identical with an internal conserved region. Based on this evidence, we postulate that the 29.5-kD polypeptide is an internal CA in C. reinhardtii and that the enzyme is closely related to the alpha-type CAs observed in animal species.

Place, publisher, year, edition, pages
1995. Vol. 109, no 2, 533-539 p.
URN: urn:nbn:se:umu:diva-34525DOI: 10.1104/pp.109.2.533PubMedID: 7480345OAI: diva2:322708
Available from: 2010-06-08 Created: 2010-06-08 Last updated: 2015-04-29Bibliographically approved

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Karlsson, JanHiltonen, ThomasSamuelsson, Göran
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Department of Plant PhysiologyUmeå Plant Science Centre (UPSC)
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