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Characterization of the pyrenoid isolated from unicellular green algaChlamydomonas reinhardtii: Particulate form of RuBisCO protein

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Summary

The pyrenoid is a protein complex in the chloroplast stroma of eukaryotic algae. After the treatment with mercury chloride, pyrenoids were isolated by sucrose density gradient centrifugation from cell-wall less mutant cells, CW-15, as well as wild type cells, C-9, of unicellular green algaChlamydomonas reinhardtii. Pyrenoids were characterized as a fraction whose protein/chlorophyll ratio was very high, and also examined by Nomarski differential interference microscopy. Most of the components consisted of 55 kDa and 16 kDa polypeptides (1∶1) which were immunologically identified as the large and small subunit of RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) protein, respectively. Some minor polypeptides were also detected. Substantial amount of RuBisCO protein is present as a particulate form in the pyrenoid in addition to the soluble form in algal chloroplast stroma.

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Abbreviations

BPB:

bromophenol blue

DAB:

3,3′-diaminobenzidine

DTT:

dithiothreitol

ELISA:

enzyme-linked immunosorbent assay

High-CO2 cells:

cells grown under air enriched with 4% CO2

Low-CO2 cells:

cells grown under ordinary air (containing 0.04% CO2)

NP-40:

nonionic detergent (Nonidet) P-40

PAGE:

polyacrylamide gel electrophoresis

PAP:

peroxidase-antiperoxidase conjugate

RuBisCO:

ribulose-1,5-bisphosphate carboxylase/oxygenase

RuBP:

ribulose-1,5-bisphosphate

SDS:

sodium dodecylsulfate

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Authors and Affiliations

  1. Institute of Applied Microbiology, University of Tokyo, Japan

    K. Kuchitsu, M. Tsuzuki & S. Miyachi

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  1. K. Kuchitsu
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  2. M. Tsuzuki
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  3. S. Miyachi
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Kuchitsu, K., Tsuzuki, M. & Miyachi, S. Characterization of the pyrenoid isolated from unicellular green algaChlamydomonas reinhardtii: Particulate form of RuBisCO protein. Protoplasma 144, 17–24 (1988). https://doi.org/10.1007/BF01320276

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  • DOI: https://doi.org/10.1007/BF01320276

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