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Analysis for Protein Glycosylation of Pattern Recognition Receptors in Plants

  • Takaakira Inokuchi
  • Yusuke SaijoEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1578)

Abstract

Recognition of molecules typical of microbes or aberrant cellular states, termed microbe- or danger-associated molecular patterns (MAMPs/DAMPs), respectively, provides an important step in plant and animal innate immunity. In plants, pattern recognition receptors (PRRs) identified to date are limited to membrane-associated proteins, of which the majority has an extracellular leucine-rich repeat (LRR) or lysine-motif (LysM) domain. These PRRs undergo quality control (QC) in the Endoplasmic Reticulum (ER) that is dependent on Asn (N)-linked glycosylation (Glc3Man9GlcNAc2 conjugation) of their extracellular domain. In Arabidopsis, genetic studies have revealed that a subset of these PRRs require an intact N-glycosylation pathway in the ER for their biogenesis and function. Here, we describe methods for immunoblot-based detection of protein glycosylation states in plants.

Key words

Plant immunity Endoplasmic Reticulum quality control (ERQC) Pattern recognition receptors (PRRs) Immunoblot analysis N-linked glycosylation Glycosidase treatment 

Notes

Acknowledgments

The researches in the Saijo Lab have been supported in part by JST PRESTO and Grant-in-Aid for Scientific Research to Y.S.

References

  1. 1.
    Saijo Y (2010) ER quality control of immune receptors and regulators in plants. Cell Microbiol 12:716–724CrossRefPubMedGoogle Scholar
  2. 2.
    Saijo Y, Tintor N, Lu X, Rauf P, Pajerowska-Mukhtar K, Haweker H, Dong X, Robatzek S, Schulze-Lefert P (2009) Receptor quality control in the endoplasmic reticulum for plant innate immunity. EMBO J 28:3439–3449CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Graduate School of Biological SciencesNara Institute of Science and TechnologyIkomaJapan
  2. 2.Japan Science and Technology Agency (JST)Precursory Research for Embryonic Science and Technology (PRESTO)KawaguchiJapan

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