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

Research output: Contribution to journalJournal articlepeer-review

  • Takayuki Ohnuma
  • Tomoyuki Numata
  • Takuo Osawa
  • Mamiko Mizuhara
  • Outi Lampela
  • André H Juffer
  • Skriver, Karen
  • Tamo Fukamizo
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 (ß/a)(8) fold with a small insertion domain composed of an a-helix and a five-stranded ß-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
JournalPlanta
Volume234
Issue number1
Pages (from-to)123-37
Number of pages15
ISSN0032-0935
DOIs
Publication statusPublished - 2011

    Research areas

  • Abscisic Acid, Amino Acid Sequence, Arabidopsis, Chitinase, Crystallography, X-Ray, Cyclopentanes, Flagellin, Gene Expression Regulation, Plant, Genes, Plant, Molecular Sequence Data, Osmosis, Oxylipins, Plant Growth Regulators, Sodium Chloride

ID: 37660699