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Chloroplast Research in Arabidopsis pp 19–30Cite as

Analysis of Thylakoid Protein Complexes by Two-Dimensional Electrophoretic Systems

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

Abstract

Photosynthetic machinery in the thylakoid membrane is prone to modifications depending on ­environmental, developmental, and morphological parameters. Such plasticity in the composition of the thylakoid membrane protein complexes guarantees efficient function of the photosynthetic machinery. In this chapter, we describe methods for separation of thylakoid membrane protein complexes at high resolution by two-dimensional gel electrophoretic systems. Solubilization of the thylakoid membrane protein complexes either by dodecylmaltoside or digitonin is described first. Then, two partially overlapping ­methods, blue native gel electrophoresis and high-resolution clear native gel electrophoresis, are demonstrated to separate the individual protein complexes. Finally, denaturing SDS-polyacrylamide gel electrophoresis is used to reveal the protein composition of each complex. Critical points in all protocols are addressed and representative examples of the composition of Arabidopsis thaliana thylakoid membrane protein complexes are shown.

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References

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Authors and Affiliations

  1. Molecular Plant Biology, Department of Biochemistry and Food Chemistry, University of Turku, Turku, Finland

    Sari Sirpiö, Marjaana Suorsa & Eva-Mari Aro

Authors
  1. Sari Sirpiö
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  2. Marjaana Suorsa
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  3. Eva-Mari Aro
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Corresponding author

Correspondence to Eva-Mari Aro .

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Editors and Affiliations

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

    R. Paul Jarvis

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Sirpiö, S., Suorsa, M., Aro, EM. (2011). Analysis of Thylakoid Protein Complexes by Two-Dimensional Electrophoretic Systems. 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_2

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  • DOI: https://doi.org/10.1007/978-1-61779-237-3_2

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-236-6

  • Online ISBN: 978-1-61779-237-3

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