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Multiple sources of carbonic anhydrase activity in pea thylakoids: soluble and membrane-bound forms

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Abstract

Carbonic anhydrase (CA) activity of pea thylakoids, thylakoid membranes enriched with photosystem I (PSI-membranes), or photosystem II (PSII-membranes) as well as both supernatant and pellet after precipitation of thylakoids treated with detergent Triton X-100 were studied. CA activity of thylakoids in the presence of varying concentrations of Triton X-100 had two maxima, at Triton/chlorophyll (triton/Chl) ratios of 0.3 and 1.0. CA activities of PSI-membranes and PSII-membranes had only one maximum each, at Triton/Chl ratio 0.3 or 1.0, respectively. Two CAs with characteristics of the membrane-bound proteins and one CA with characteristics of the soluble proteins were found in the medium after thylakoids were incubated with Triton. One of the first two CAs had mobility in PAAG after native electrophoresis the same as that of CA residing in PSI-membranes, and the other CA had mobility the same as the mobility of CA residing in PSII-membranes, but the latter was different from CA situated in PSII core-complex (Ignatova et al. 2006 Biochemistry (Moscow) 71:525–532). The properties of the “soluble” CA removed from thylakoids were different from the properties of the known soluble CAs of plant cell: apparent molecular mass was about 262 kD and it was three orders more sensitive to the specific CA inhibitor, ethoxyzolamide, than soluble stromal CA. The data are discussed as indicating the presence of, at least, four CAs in pea thylakoids.

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Abbreviations

AA:

Acetazolamide

CA:

Carbonic anhydrase

DM:

Dodecyl-β-D-maltoside

DTT:

1,4-Dithio-DL-threitol

EZ:

Ethoxyzolamide

PAAG:

Polyacrylamide gel

PMSF:

Phenylmethylsulfonylfluoride

PSI:

Photosystem I

PSII:

Photosystem II

Rubisco:

Ribulosebisphosphatecarboxylase/oxygenase

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Acknowledgments

The authors express their gratitude to Dr. M.S. Khristin for valuable discussions.

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Authors and Affiliations

  1. Photosynthetic Electron Transport Laboratory, Institute of Basic Biological Problems, Russian Academy of Sciences, Institutskaya, 2, Pushchino Moscow Region, 142290, Russian Federation

    Natalia N. Rudenko, Lyudmila K. Ignatova & Boris N. Ivanov

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  1. Natalia N. Rudenko
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  2. Lyudmila K. Ignatova
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  3. Boris N. Ivanov
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Correspondence to Boris N. Ivanov.

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Rudenko, N.N., Ignatova, L.K. & Ivanov, B.N. Multiple sources of carbonic anhydrase activity in pea thylakoids: soluble and membrane-bound forms. Photosynth Res 91, 81–89 (2007). https://doi.org/10.1007/s11120-007-9148-2

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