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Characterization of Proteins Localized to Plant ER-PM Contact Sites

Wang, Pengwei; Hawes, Chris; Richardson, Christine; Hussey, Patrick J.

Authors

Pengwei Wang

Chris Hawes

Christine Richardson



Contributors

Chris Hawes
Editor

Verena Kriechbaumer
Editor

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.

Citation

Wang, P., Hawes, C., Richardson, C., & Hussey, P. J. (2018). Characterization of Proteins Localized to Plant ER-PM Contact Sites. In C. Hawes, & V. Kriechbaumer (Eds.), The Plant Endoplasmic Reticulum (23-31). Humana Press. https://doi.org/10.1007/978-1-4939-7389-7_3

Online Publication Date Oct 17, 2017
Publication Date 2018
Deposit Date Oct 18, 2017
Publisher Humana Press
Pages 23-31
Series Title Methods in Molecular Biology
Series Number 1691
Book Title The Plant Endoplasmic Reticulum
ISBN 9781493973880
DOI https://doi.org/10.1007/978-1-4939-7389-7_3