A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis
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)...
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| Veröffentlicht in: | Planta 2011-07, Vol.234 (1), p.123-137 |
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| creator | Ohnuma, Takayuki Numata, Tomoyuki Osawa, Takuo Mizuhara, Mamiko Lampela, Outi Juffer, André H. Skriver, Karen Fukamizo, Tamo |
| description | 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. |
| doi_str_mv | 10.1007/s00425-011-1390-3 |
| format | Article |
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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.</description><identifier>ISSN: 0032-0935</identifier><identifier>EISSN: 1432-2048</identifier><identifier>DOI: 10.1007/s00425-011-1390-3</identifier><identifier>PMID: 21390509</identifier><identifier>CODEN: PLANAB</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer</publisher><subject>Abscisic Acid - adverse effects ; Agriculture ; Amino Acid Sequence ; Amino acids ; Arabidopsis - enzymology ; Arabidopsis - genetics ; Arabidopsis - metabolism ; Biological and medical sciences ; Biomedical and Life Sciences ; Chitinases - chemistry ; Chitinases - genetics ; Chitinases - metabolism ; Crystal structure ; Crystallography, X-Ray ; Cyclopentanes - adverse effects ; E coli ; Ecology ; Enzymes ; Flagellin - adverse effects ; Forestry ; Fundamental and applied biological sciences. 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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.</description><subject>Abscisic Acid - adverse effects</subject><subject>Agriculture</subject><subject>Amino Acid Sequence</subject><subject>Amino acids</subject><subject>Arabidopsis - enzymology</subject><subject>Arabidopsis - genetics</subject><subject>Arabidopsis - metabolism</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Chitinases - chemistry</subject><subject>Chitinases - genetics</subject><subject>Chitinases - metabolism</subject><subject>Crystal structure</subject><subject>Crystallography, X-Ray</subject><subject>Cyclopentanes - adverse effects</subject><subject>E coli</subject><subject>Ecology</subject><subject>Enzymes</subject><subject>Flagellin - adverse effects</subject><subject>Forestry</subject><subject>Fundamental and applied biological sciences. 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Psychology</topic><topic>Gene expression</topic><topic>Gene Expression Regulation, Plant</topic><topic>Genes, Plant - physiology</topic><topic>Hormones</topic><topic>Hydrogen</topic><topic>Hydrogen bonds</topic><topic>Life Sciences</topic><topic>Ligands</topic><topic>Molecular Sequence Data</topic><topic>Oligosaccharides</topic><topic>Original Article</topic><topic>Osmosis</topic><topic>Osmosis - physiology</topic><topic>Oxylipins - adverse effects</topic><topic>Plant Growth Regulators - metabolism</topic><topic>Plant Sciences</topic><topic>Plants</topic><topic>Sodium</topic><topic>Sodium chloride</topic><topic>Sodium Chloride - adverse effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ohnuma, Takayuki</creatorcontrib><creatorcontrib>Numata, Tomoyuki</creatorcontrib><creatorcontrib>Osawa, Takuo</creatorcontrib><creatorcontrib>Mizuhara, Mamiko</creatorcontrib><creatorcontrib>Lampela, Outi</creatorcontrib><creatorcontrib>Juffer, André H.</creatorcontrib><creatorcontrib>Skriver, Karen</creatorcontrib><creatorcontrib>Fukamizo, Tamo</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><jtitle>Planta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ohnuma, Takayuki</au><au>Numata, Tomoyuki</au><au>Osawa, Takuo</au><au>Mizuhara, Mamiko</au><au>Lampela, Outi</au><au>Juffer, André H.</au><au>Skriver, Karen</au><au>Fukamizo, Tamo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis</atitle><jtitle>Planta</jtitle><stitle>Planta</stitle><addtitle>Planta</addtitle><date>2011-07-01</date><risdate>2011</risdate><volume>234</volume><issue>1</issue><spage>123</spage><epage>137</epage><pages>123-137</pages><issn>0032-0935</issn><eissn>1432-2048</eissn><coden>PLANAB</coden><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.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer</pub><pmid>21390509</pmid><doi>10.1007/s00425-011-1390-3</doi><tpages>15</tpages></addata></record> |
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| subjects | Abscisic Acid - adverse effects Agriculture Amino Acid Sequence Amino acids Arabidopsis - enzymology Arabidopsis - genetics Arabidopsis - metabolism Biological and medical sciences Biomedical and Life Sciences Chitinases - chemistry Chitinases - genetics Chitinases - metabolism Crystal structure Crystallography, X-Ray Cyclopentanes - adverse effects E coli Ecology Enzymes Flagellin - adverse effects Forestry Fundamental and applied biological sciences. Psychology Gene expression Gene Expression Regulation, Plant Genes, Plant - physiology Hormones Hydrogen Hydrogen bonds Life Sciences Ligands Molecular Sequence Data Oligosaccharides Original Article Osmosis Osmosis - physiology Oxylipins - adverse effects Plant Growth Regulators - metabolism Plant Sciences Plants Sodium Sodium chloride Sodium Chloride - adverse effects |
| title | A class V chitinase from Arabidopsis thaliana: gene responses, enzymatic properties, and crystallographic analysis |
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