Chapter title |
In Vivo Radiolabeling of Arabidopsis Chloroplast Proteins and Separation of Thylakoid Membrane Complexes by Blue Native PAGE
|
---|---|
Chapter number | 19 |
Book title |
Plant Proteostasis
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3759-2_19 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3757-8, 978-1-4939-3759-2
|
Authors |
Catharina Nickel, Thomas Brylok, Serena Schwenkert |
Editors |
L. Maria Lois, Rune Matthiesen |
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 (35)S-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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