A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis
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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, André H; Skriver, Karen; Fukamizo, Tamo.
In: Planta, Vol. 234, No. 1, 2011, p. 123-37.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis
AU - Ohnuma, Takayuki
AU - Numata, Tomoyuki
AU - Osawa, Takuo
AU - Mizuhara, Mamiko
AU - Lampela, Outi
AU - Juffer, André H
AU - Skriver, Karen
AU - Fukamizo, Tamo
PY - 2011
Y1 - 2011
N2 - 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.
AB - 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.
KW - Abscisic Acid
KW - Amino Acid Sequence
KW - Arabidopsis
KW - Chitinase
KW - Crystallography, X-Ray
KW - Cyclopentanes
KW - Flagellin
KW - Gene Expression Regulation, Plant
KW - Genes, Plant
KW - Molecular Sequence Data
KW - Osmosis
KW - Oxylipins
KW - Plant Growth Regulators
KW - Sodium Chloride
U2 - 10.1007/s00425-011-1390-3
DO - 10.1007/s00425-011-1390-3
M3 - Journal article
C2 - 21390509
VL - 234
SP - 123
EP - 137
JO - Planta
JF - Planta
SN - 0032-0935
IS - 1
ER -
ID: 37660699