Protoplasma

, Volume 144, Issue 1, pp 17–24 | Cite as

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

  • K. Kuchitsu
  • M. Tsuzuki
  • S. Miyachi
Article

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.

Keywords

Chlamydomonas Pyrenoid Ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) Stroma 

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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Copyright information

© Springer-Verlag 1988

Authors and Affiliations

  • K. Kuchitsu
    • 1
  • M. Tsuzuki
    • 1
  • S. Miyachi
    • 1
  1. 1.Institute of Applied MicrobiologyUniversity of TokyoJapan

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