Abstract
The investigation of membrane protein complex assembly and degradation is essential to understand cellular protein dynamics. Blue native PAGE provides a powerful tool to analyze the composition and formation of protein complexes. Combined with in vivo radiolabeling, the synthesis and decay of protein complexes can be monitored on a timescale ranging from minutes to several hours. Here, we describe a protocol to analyze thylakoid membrane complexes starting either with 35S-methionine labeling of intact Arabidopsis leaves to investigate protein complex dynamics or with unlabeled leaf material to monitor steady-state complex composition.
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Acknowledgements
Financial support from the German Research Council (DFG, SFB1035, project A4, to SS) is acknowledged. We would further like to thank Manuela Urbischek for providing figures as well as Peter Hagl for critical reading of the manuscript.
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Nickel, C., Brylok, T., Schwenkert, S. (2016). In Vivo Radiolabeling of Arabidopsis Chloroplast Proteins and Separation of Thylakoid Membrane Complexes by Blue Native PAGE. In: Lois, L., Matthiesen, R. (eds) Plant Proteostasis. Methods in Molecular Biology, vol 1450. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3759-2_19
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DOI: https://doi.org/10.1007/978-1-4939-3759-2_19
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