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TECHNICAL ADVANCE: Functional immobilization of plant receptor‐like kinase onto microbeads towards receptor array construction and receptor‐based ligand fishing
Author(s) -
Shinohara Hidefumi,
Matsubayashi Yoshikatsu
Publication year - 2007
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2007.03204.x
Subject(s) - ligand (biochemistry) , receptor , biology , microbead (research) , kinase , confocal laser scanning microscopy , confocal , biophysics , biochemistry , combinatorial chemistry , chemistry , physics , optics
Summary Despite the large number of receptor‐like kinases (RLKs) in plants, few of their specific ligands are known. Ligand fishing is one of the most challenging post‐genomic technologies. Here, we report a strategy for functional immobilization of plant RLKs on microbeads via covalent linkage. Because of the high density of immobilized RLKs, ligand‐receptor interaction can be visualized at high sensitivity using fluorescent‐labeled ligands under the confocal laser scanning microscope. Moreover, using a receptor‐based affinity chromatography system with RLK microbeads, the ligand of the receptor was directly retrieved at high purity from complex natural samples. Our RLK microbeads and receptor‐based ligand fishing approach is a feasible alternative to conventional forward genetics and bioassay‐based biochemical purification for identification of novel ligand–receptor pairs in plants.