Affinity Labeling and Purification of Plant Chitin-Binding LysM Receptor with Chitin Octasaccharide Derivatives

Tomonori Shinya, Naoto Shibuya, Hanae Kaku

Research output: Contribution to journalArticle

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
Pages (from-to)401-412
Number of pages12
JournalMethods in molecular biology (Clifton, N.J.)
Volume2132
DOIs
Publication statusPublished - 1 Jan 2020

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Keywords

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

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