Abstract
Mass-spectrometric measurements of 18O exchange from 13C18O2 were used to follow changes in the intracellular carbonic anhydrase (CA) activity of cells of Chlamydomonas reinhardtii Dang, wild type and the ca-1 mutant during adaptation to air. With intact cells as well as with crude homogenates total intracellular CA activity in wild-type cells increased six to tenfold within 4 h after transferring cells from 5% CO2 (high inorganic carbon, Ci) to ambient air (air adapted). After that time the activity slowly declined to a level similar to that observed with cells which had been continuously grown in air (low-Ci grown). In the ca-1 mutant, total CA was induced to a similar extent during 4 h of adaptation; however, absolute activities were two to three times lower in ca-1 than in the wild type regardless of the CO2 supply. When crude extracts from wild-type cells were separated into soluble and insoluble fractions, each fraction contained about half of the internal CA activity. Within 4 h of adaptation, both forms of CA activity were simultaneously enhanced by nine to tenfold, reaching levels similar to those found in low-Cigrown cells. In contrast, in the ca-1 mutant the soluble CA activity was only enhanced by about eightfold while the level of insoluble CA was very low even in low-Ci cells. After isolation of intact chloroplasts from wild-type cells and further subfractionation, around 70–80% of total chloroplastic CA activity was found to be in the insoluble fraction while 17–20% remained in the soluble fraction. Both chloroplastic CA activities were inducible within the first 4 h of adaptation to air, with each of them being eight to ten times higher than in high-Ci algae. After that time their activities were similar to the corresponding CA values in low-Ci-grown cells. In contrast, plastids from high-Ci cells of the ca-1 mutant showed 40% less insoluble-CA activity compared to the wild type and this insoluble-CA activity was not increased at all by transferring algae to air. In addition, no soluble-CA activity was detected in chloroplasts from high-Ci and air-adapted ca-1 cells. These results indicate the presence of three intracellular CA activities in high-Ci air-adapted and low-Ci cells of the wild type and that two of them are associated with the chloroplasts. All three activities are completely induced within the first 4 h of adaptation to air in wild-type cells. In contrast, it was not possible to induce any of the chloroplastic CA activities in the ca-1 mutant. The possibility that the soluble chloroplastic CA represents a pyrenoid-located CA is discussed.
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Abbreviations
- AZA:
-
acetazolamide
- BTP:
-
Bis-tris-propane (1,3-bis[tris(hydroxymethyl)methylamino]propane)
- Ci :
-
inorganic carbon (HCO −3 +CO2)
- CA:
-
carbonic anhydrase
- cCAsol :
-
insoluble carbonic anhydrase activity
- Chl:
-
chlorophyll
- CCM:
-
CO2-concentrating mechanism
- high-Ci cells:
-
cells grown in air enriched with 5% CO2
- low-Ci cells:
-
cells grown in air (0.035% CO2)
- Rubisco:
-
ribulose1,5-bisphosphate carboxylase/oxygenase
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This work is dedicated to Professor A. Wild on the occasion of his 65th birthday
We are grateful to Drs. Murray Badger, Dean Price and Jian-Wei Yu, (Molecular Plant Physiology Group, Research School of Biological Sciences, The Australian National University, Canberra, Australia) for critically reading the manuscript. We also wish to thank Drs. Martin Spalding and Laura Marek (Dept. of Botany, Iowa State University, Ames, Iowa, USA) for providing us with the ca-1 mutant. This work was supported by a grant from the Deutsche Forschungsgemeinschaft to H.P.F.
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Sültemeyer, D., Amoroso, G. & Fock, H. Induction of intracellular carbonic anhydrases during the adaptation to low inorganic carbon concentrations in wild-type and ca-1 mutant cells of Chlamydomonas reinhardtii . Planta 196, 217–224 (1995). https://doi.org/10.1007/BF00201377
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DOI: https://doi.org/10.1007/BF00201377