Functional immobilization of plant receptor-like kinase onto microbeads towards receptor array construction and receptor-based ligand fishing.
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.
Graduate School of Bio-Agricultural Sciences, Nagoya University, Chikusa, Nagoya 464-8601, Japan.
Hidefumi
H
Shinohara
Yoshikatsu
Y
Matsubayashi
D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid; D017931:DNA Primers; D005456:Fluorescent Dyes; D008024:Ligands; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D010940:Plant Proteins / Q000378:metabolism*; D010944:Plants / Q000201:enzymology*; D011494:Protein Kinases / Q000378:metabolism*
D017931:DNA Primers; D005456:Fluorescent Dyes; D008024:Ligands; D010940:Plant Proteins; D011494:Protein Kinases