Affinity labeling and purification of plant chitin-binding LysM receptor with chitin octasaccharide derivatives

Tomonori Shinya, Naoto Shibuya, Hanae Kaku

    Research output: Chapter in Book/Report/Conference proceedingChapter

    Abstract

    Lysin motif (LysM) is a carbohydrate-binding modules found in all kingdoms. LysM binds to N-acetylglucosamine-containing molecules such as peptidoglycan, chitin, Nod factor, and Myc factor and is found in peptidoglycan hydrolases, chitinases, and plant pathogen effectors and plant receptor/co-receptor for defense and symbiosis signaling. This chapter describes the synthesis of a nonradioactive chitin ligand, biotinylated chitin octasaccharide, (GlcNAc)8-Bio, and its application for the detection and characterization of chitin-binding LysM receptor CEBiP in the microsomal membrane fraction of rice suspension-cultured cells by affinity labeling. We also describe the purification of CEBiP from the plasma membrane of the rice cells by affinity chromatography with the synthesized (GlcNAc)8-APEA-CH-Sepharose as an affinity matrix.

    Original languageEnglish
    Title of host publicationMethods in Molecular Biology
    PublisherHumana Press Inc.
    Pages401-412
    Number of pages12
    DOIs
    Publication statusPublished - 2020

    Publication series

    NameMethods in Molecular Biology
    Volume2132
    ISSN (Print)1064-3745
    ISSN (Electronic)1940-6029

    Keywords

    • Affinity labeling
    • Affinity ligand
    • Biotinylation
    • CEBiP
    • Chitin oligosaccharide
    • Chitin receptor
    • LysM
    • Purification

    ASJC Scopus subject areas

    • Molecular Biology
    • Genetics

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