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Preparation and Proteomic Analysis of Chloroplast Ribosomes

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Book cover Chloroplast Research in Arabidopsis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 775))

Abstract

Proteomics of chloroplast ribosomes in spinach and Chlamydomonas revealed unique protein composition and structures of plastid ribosomes. These studies have suggested the presence of some ribosomal proteins unique to plastid ribosomes which may be involved in plastid-unique translation regulation. Considering the strong background of genetic analysis and molecular biology in Arabidopsis, the in-depth proteomic characterization of Arabidopsis plastid ribosomes would facilitate further understanding of plastid translation in higher plants. Here, I describe simple and rapid methods for the preparation of plastid ribosomes from Chlamydomonas and Arabidopsis using sucrose gradients. I also describe purity criteria and methods for yield estimation of the purified plastid ribosomes and subunits, methods for the preparation of plastid ribosomal proteins, as well as the identification of some Arabidopsis plastid ribosomal proteins by matrix-assisted laser desorption/ionization mass spectrometry.

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Acknowledgments

I thank Dr. Alap R. Subramanian, Dr. Stephen P. Mayfield, Dr. Don P. Bourque, and Dr. Tatsuya Oda for their support and advice. I am grateful to Dr. Paul Jarvis for his encouragement and helpful comments on the manuscript. I also thank Ms. Sawako Iwanaga, Ms. Moemi Yamawaki, and Ms. Akiko Myouse for technical assistance with plant/algal culture and ribosome preparation. This work was supported by funds from the Grant-in-Aid from the Japan Society for the Promotion of Science (16770036), and in part by a Grant-in-Aid for Scientific Research from Nagasaki University, Japan.

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  1. Division of Biochemistry, Nagasaki University, Nagasaki, Japan

    Kenichi Yamaguchi

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  1. Kenichi Yamaguchi
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Correspondence to Kenichi Yamaguchi .

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  1. , Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, United Kingdom

    R. Paul Jarvis

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Yamaguchi, K. (2011). Preparation and Proteomic Analysis of Chloroplast Ribosomes. In: Jarvis, R. (eds) Chloroplast Research in Arabidopsis. Methods in Molecular Biology, vol 775. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-237-3_13

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  • DOI: https://doi.org/10.1007/978-1-61779-237-3_13

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  • Publisher Name: Humana Press, Totowa, NJ

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