Chapter title |
Characterization of Proteins Localized to Plant ER-PM Contact Sites
|
---|---|
Chapter number | 3 |
Book title |
The Plant Endoplasmic Reticulum
|
Published in |
Methods in molecular biology, January 2018
|
DOI | 10.1007/978-1-4939-7389-7_3 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7388-0, 978-1-4939-7389-7
|
Authors |
Pengwei Wang, Chris Hawes, Christine Richardson, Patrick J. Hussey |
Abstract |
Like in most eukaryotic cells, the plant endoplasmic reticulum (ER) network is physically linked to the plasma membrane (PM), forming ER-PM contact sites (EPCS). The protein complex required for maintaining the EPCS is composed of ER integral membrane proteins (e.g., VAP27, synaptotagmins), PM-associated proteins (e.g., NET3C), and the cytoskeleton. Here, we describe methods for identifying possible EPCS-associated proteins. These include GFP-tagged protein expression followed by image analysis, and immuno-gold labeling at the ultrastructural level. In combination, these methods can be used to identify the localization of putative EPCS proteins as well as used to postulate their subcellular function. |
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