| RRC ID |
12636
|
| 著者 |
Iizasa E, Mitsutomi M, Nagano Y.
|
| タイトル |
Direct binding of a plant LysM receptor-like kinase, LysM RLK1/CERK1, to chitin in vitro.
|
| ジャーナル |
J Biol Chem
|
| Abstract |
Plants induce immune responses against fungal pathogens by recognition of chitin, which is a component of the fungal cell wall. Recent studies have revealed that LysM receptor-like kinase 1/chitin elicitor receptor kinase 1 (LysM RLK1/CERK1) is a critical component for the immune responses to chitin in Arabidopsis thaliana. However, the molecular mechanism of the chitin recognition by LysM RLK1 still remains unknown. Here, we present the first evidence for direct binding of LysM RLK1 to chitin. We expressed LysM RLK1 fused with yeast-enhanced green fluorescent protein (LysM RLK1-yEGFP) in yeast cells. Binding studies using the solubilized LysM RLK1-yEGFP and several insoluble polysaccharides having similar structures showed that LysM RLK1-yEGFP specifically binds to chitin. Subsequently, the fluorescence microscopic observation of the solubilized LysM RLK1-yEGFP binding to chitin beads revealed that the binding was saturable and had a high affinity, with a K(d) of approximately 82 nm. This binding was competed by the addition of soluble glycol chitin or high concentration of chitin oligosaccharides having 4-8 residues of N-acetyl glucosamine. However, the competition of these chitin oligosaccharides is weaker than that of glycol chitin. These data suggest that LysM RLK1 has a higher affinity for chitin having a longer residue of N-acetyl glucosamine. We also found that LysM RLK1-yEGFP was autophosphorylated in vitro and that chitin does not affect the phosphorylation of LysM RLK1-yEGFP. Our results provide a new dimension to chitin elicitor perception in plants.
|
| 巻・号 |
285(5)
|
| ページ |
2996-3004
|
| 公開日 |
2010-1-29
|
| DOI |
10.1074/jbc.M109.027540
|
| PII |
S0021-9258(20)64794-9
|
| PMID |
19951949
|
| PMC |
PMC2823440
|
| MeSH |
Acetylglucosamine / chemistry
Amino Acid Motifs
Arabidopsis / metabolism
Arabidopsis Proteins / metabolism*
Chitin / chemistry*
Cloning, Molecular
Green Fluorescent Proteins / chemistry
Inhibitory Concentration 50
Kinetics
Ligands
Microscopy, Fluorescence / methods
Phosphorylation
Plant Proteins / metabolism*
Protein Binding
Protein Serine-Threonine Kinases / metabolism*
Saccharomyces cerevisiae / metabolism
|
| IF |
4.238
|
| 引用数 |
119
|
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| リソース情報 |
| 酵母 |
BY2777 |
