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A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis

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Expression of a class V chitinase gene (At4g19810, AtChiC) in Arabidopsis thaliana was examined by quantitative real-time PCR and by analyzing microarray data available at Genevestigator. The gene expression was induced by the plant stress-related hormones abscisic acid (ABA) and jasmonic acid (JA) and by the stress resulting from the elicitor flagellin, NaCl, and osmosis. The recombinant AtChiC protein was produced in E. coli, purified, and characterized with respect to the structure and function. The recombinant AtChiC hydrolyzed N-acetylglucosamine oligomers producing dimers from the non-reducing end of the substrates. The crystal structure of AtChiC was determined by the molecular replacement method at 2.0 Å resolution. AtChiC was found to adopt an (β/α)8 fold with a small insertion domain composed of an α-helix and a five-stranded β-sheet. From docking simulation of AtChiC with pentameric substrate, the amino acid residues responsible for substrate binding were found to be well conserved when compared with those of the class V chitinase from Nicotiana tabacum (NtChiV). All of the structural and functional properties of AtChiC are quite similar to those obtained for NtChiV, and seem to be common to class V chitinases from higher plants.

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Class V chitinase from Arabidopsis thaliana


Class V chitinase from Nicotiana tabacum


Family GH-18 chitinase B from Serratia marcescens



(GlcNAc) n :

β-1,4-Linked oligosaccharide of GlcNAc with a polymerization degree of n


Polymerase chain reaction


Sodium dodecyl sulfate-polyacrylamide gel electrophoresis


High performance liquid chromatography


Matrix-assisted laser desorption ionization time-of-flight mass spectrometry


Abscisic acid


Jasmonic acid


Gibberellic acid


Root mean square


Expression sequence tag


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This work was partly supported by a grant to O.L. from the North Ostrobothnia Regional Fund of the Finnish Cultural Foundation. The authors thank the beam-line staffs at BL-17A of KEK (Ibaraki, Japan) for technical assistance during data collection, and Hideko Inanaga of AIST for technical assistance on protein crystallography.

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Correspondence to Tomoyuki Numata or Tamo Fukamizo.

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Ohnuma, T., Numata, T., Osawa, T. et al. A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis. Planta 234, 123–137 (2011).

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