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. 1978 Jul;62(1):97–100. doi: 10.1104/pp.62.1.97

Biosynthesis of Ribulose-1,5-Bisphosphate Carboxylase in Spinach Leaf Protoplasts 1

Mikio Nishimura 1, Takashi Akazawa 1
PMCID: PMC1092063  PMID: 16660478

Abstract

Spinach leaf (Spinacia oleracea L. var. Kyoho) protoplasts sustain protein-synthesizing activity as measured by the incorporation of [14C]-leucine into the protein fraction both in the light and in the dark. By the immunoprecipitation of ribulose-1,5-bisphosphate (RuP2) carboxylase with rabbit antibody raised against the purified spinach enzyme preparation, it was found that approximately 7% of the total radiocarbon incorporated into the protein fraction in the light was in the carboxylase molecules. However, there was no measurable net increase observed in the content of the enzyme protein in the experimental conditions employed. It was found that both chloramphenicol and cycloheximide inhibited the incorporation of [14C]leucine into RuP2 carboxylase and its constituent subunits, as measured by the immunoprecipitation of the enzyme molecule and its subunits, A and B.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Blair G. E., Ellis R. J. Protein synthesis in chloroplasts. I. Light-driven synthesis of the large subunit of fraction I protein by isolated pea chloroplasts. Biochim Biophys Acta. 1973 Aug 24;319(2):223–234. doi: 10.1016/0005-2787(73)90013-0. [DOI] [PubMed] [Google Scholar]
  2. Cashmore A. R. Protein synthesis in plant leaf tissue. The sites of synthesis of the major proteins. J Biol Chem. 1976 May 10;251(9):2848–2853. [PubMed] [Google Scholar]
  3. Criddle R. S., Dau B., Kleinkopf G. E., Huffaker R. C. Differential synthesis of ribulosediphosphate carboxylase subunits. Biochem Biophys Res Commun. 1970 Nov 9;41(3):621–627. doi: 10.1016/0006-291x(70)90058-6. [DOI] [PubMed] [Google Scholar]
  4. Dobberstein B., Blobel G., Chua N. H. In vitro synthesis and processing of a putative precursor for the small subunit of ribulose-1,5-bisphosphate carboxylase of Chlamydomonas reinhardtii. Proc Natl Acad Sci U S A. 1977 Mar;74(3):1082–1085. doi: 10.1073/pnas.74.3.1082. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gooding L. R., Roy H., Jagendorf A. T. Immunological identification of nascent subunits of wheat ribulose diphosphate carboxylase on ribosomes of both chloroplast and cytoplasmic origin. Arch Biochem Biophys. 1973 Nov;159(1):324–335. doi: 10.1016/0003-9861(73)90458-x. [DOI] [PubMed] [Google Scholar]
  6. Hartley M. R., Wheeler A., Ellis R. J. Protein synthesis in chloroplasts. V. Translation of messenger RNA for the large subunit of fraction I protein in a heterologous cell-free system. J Mol Biol. 1975 Jan 5;91(1):67–77. doi: 10.1016/0022-2836(75)90372-1. [DOI] [PubMed] [Google Scholar]
  7. Iwanij V., Chua N. H., Siekevitz P. Synthesis and turnover of ribulose biphosphate carboxylase and of its subunits during the cell cycle of Chlamydomonas reinhardtii. J Cell Biol. 1975 Mar;64(3):572–585. doi: 10.1083/jcb.64.3.572. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. Nishimura M., Akazawa T. Photosynthetic activities of spinach leaf protoplasts. Plant Physiol. 1975 Apr;55(4):712–716. doi: 10.1104/pp.55.4.712. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nishimura M., Akazawa T. Studies on spinach leaf ribulosebisphosphate carboxylase. Carboxylase and oxygenase reaction examined by immunochemical methods. Biochemistry. 1974 May 21;13(11):2277–2281. doi: 10.1021/bi00708a006. [DOI] [PubMed] [Google Scholar]
  11. Nishimura M., Graham D., Akazawa T. Isolation of intact chloroplasts and other cell organelles from spinach leaf protoplasts. Plant Physiol. 1976 Sep;58(3):309–314. doi: 10.1104/pp.58.3.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Nishimura M., Takabe T., Sugiyama T., Akazawa T. Structure and function of chloroplast proteins. XIX. Dissociation of spinach leaf ribulose-1,5-diphosphate carboxylase by p-mercuribenzoate. J Biochem. 1973 Nov;74(5):945–954. [PubMed] [Google Scholar]
  13. Roy H., Patterson R., Jagendorf A. T. Identification of the small subunit of ribulose 1,5-bisphosphate carboxylase as a product of wheat leaf cytoplasmic ribosomes. Arch Biochem Biophys. 1976 Jan;172(1):64–73. doi: 10.1016/0003-9861(76)90048-5. [DOI] [PubMed] [Google Scholar]
  14. Roy H., Terenna B. Synthesis of the Small Subunit of Ribulose-1,5-bisphosphate Carboxylase by Soluble Fraction Polyribosomes of Pea Leaves. Plant Physiol. 1977 Oct;60(4):532–537. doi: 10.1104/pp.60.4.532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sakai F., Takebe I. RNA and protein synthesis in protoplasts isolated from tobacco leaves. Biochim Biophys Acta. 1970 Dec 14;224(2):531–540. doi: 10.1016/0005-2787(70)90585-x. [DOI] [PubMed] [Google Scholar]
  16. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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