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

Takayuki Ohnuma, Tomoyuki Numata, Takuo Osawa, Mamiko Mizuhara, Outi Lampela, A. H. Juffer, Karen Skriver, Tamo Fukamizo

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)123-137
Number of pages15
JournalPlanta
Volume234
Issue number1
DOIs
Publication statusPublished - Jul 1 2011

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Chitinases
chitinase
Arabidopsis
Arabidopsis thaliana
Genes
Osmosis
osmosis
Flagellin
flagellin
Abscisic Acid
N-acetylglucosamine
genes
Acetylglucosamine
plant stress
jasmonic acid
crystal structure
Recombinant Proteins
recombinant proteins
Nicotiana tabacum
functional properties

All Science Journal Classification (ASJC) codes

  • Genetics
  • Plant Science

Cite this

A class V chitinase from Arabidopsis thaliana : Gene responses, enzymatic properties, and crystallographic analysis. / Ohnuma, Takayuki; Numata, Tomoyuki; Osawa, Takuo; Mizuhara, Mamiko; Lampela, Outi; Juffer, A. H.; Skriver, Karen; Fukamizo, Tamo.

In: Planta, Vol. 234, No. 1, 01.07.2011, p. 123-137.

Research output: Contribution to journalArticle

Ohnuma, T, Numata, T, Osawa, T, Mizuhara, M, Lampela, O, Juffer, AH, Skriver, K & Fukamizo, T 2011, 'A class V chitinase from Arabidopsis thaliana: Gene responses, enzymatic properties, and crystallographic analysis', Planta, vol. 234, no. 1, pp. 123-137. https://doi.org/10.1007/s00425-011-1390-3
Ohnuma, Takayuki ; Numata, Tomoyuki ; Osawa, Takuo ; Mizuhara, Mamiko ; Lampela, Outi ; Juffer, A. H. ; Skriver, Karen ; Fukamizo, Tamo. / A class V chitinase from Arabidopsis thaliana : Gene responses, enzymatic properties, and crystallographic analysis. In: Planta. 2011 ; Vol. 234, No. 1. pp. 123-137.
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