+Open data
-Basic information
Entry | Database: SASBDB / ID: SASDDE7 |
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Sample | Class I chitinase 1 from Agave tequilana
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Biological species | Agave tequilana (plant) |
Citation | Journal: FEBS J / Year: 2019 Title: A biophysical and structural study of two chitinases from Agave tequilana and their potential role as defense proteins. Authors: Yusvel Sierra-Gómez / Annia Rodríguez-Hernández / Patricia Cano-Sánchez / Homero Gómez-Velasco / Alejandra Hernández-Santoyo / Dritan Siliqi / Adela Rodríguez-Romero / Abstract: Plant chitinases are enzymes that have several functions, including providing protection against pathogens. Agave tequilana is an economically important plant that is poorly studied. Here, we ...Plant chitinases are enzymes that have several functions, including providing protection against pathogens. Agave tequilana is an economically important plant that is poorly studied. Here, we identified a chitinase from short reads of the A. tequilana transcriptome (AtChi1). A second chitinase, differing by only six residues from the first, was isolated from total RNA of plants infected with Fusarium oxysporum (AtChi2). Both enzymes were overexpressed in Escherichia coli and analysis of their sequences indicated that they belong to the class I glycoside hydrolase family19, whose members exhibit two domains: a carbohydrate-binding module and a catalytic domain, connected by a flexible linker. Activity assays and thermal shift experiments demonstrated that the recombinant Agave enzymes are highly thermostable acidic endochitinases with Tm values of 75 °C and 71 °C. Both exhibit a molecular mass close to 32 kDa, as determined by MALDI-TOF, and experimental pIs of 3.7 and 3.9. Coupling small-angle x-ray scattering information with homology modeling and docking simulations allowed us to structurally characterize both chitinases, which notably show different interactions in the binding groove. Even when the six different amino acids are all exposed to solvent in the loops located near the linker and opposite to the binding site, they confer distinct kinetic parameters against colloidal chitin and similar affinity for (GlnNAc) as shown by isothermal titration calorimetry. Interestingly, binding is more enthalpy-driven for AtChi2. Whereas the physiological role of these chitinases remains unknown, we demonstrate that they exhibit important antifungal activity against chitin-rich fungi such as Aspergillus sp. DATABASE: SAXS structural data are available in the SASBDB database with accession numbers SASDDE7 and SASDDA6. ENZYMES: Chitinases (EC3.2.1.14). |
Contact author |
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-Structure visualization
Structure viewer | Molecule: MolmilJmol/JSmol |
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-Downloads & links
-Models
Model #2037 | Type: dummy / Radius of dummy atoms: 1.40 A / Chi-square value: 1.624 / P-value: 0.216071 Search similar-shape structures of this assembly by Omokage search (details) |
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-Sample
Sample | Name: Class I chitinase 1 from Agave tequilana / Specimen concentration: 0.23 mg/ml |
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Buffer | Name: MES 50 mM / pH: 6 |
Entity #1052 | Name: ChiAt1 / Type: protein / Description: Chitinase 1 / Formula weight: 31.753 / Num. of mol.: 1 / Source: Agave tequilana Sequence: GGAQQCGSQA GGAVCPNGLC CSQFGYCGST SPYCGNGCQS QCGGGSSPTP NPPSGGGGGG GGGGSGVGSI ISSSLFDQIL LHRNDAACPA HGFYTYDAFV AAANAFSGFA TTGDADTQKR EIAAFLAQTS HETTGGWPTA PDGPYSWGYC FLQEQGNPGD YCVPNDQWPC ...Sequence: GGAQQCGSQA GGAVCPNGLC CSQFGYCGST SPYCGNGCQS QCGGGSSPTP NPPSGGGGGG GGGGSGVGSI ISSSLFDQIL LHRNDAACPA HGFYTYDAFV AAANAFSGFA TTGDADTQKR EIAAFLAQTS HETTGGWPTA PDGPYSWGYC FLQEQGNPGD YCVPNDQWPC APGKKYYGRG PIQISYNYNY GPCGNAIGSD LLNNPDLVAS DPTVSFKTAL WFWMTPQSPK PSCHDVITGQ WTPSAADQAA GRVPGFGVIT NIINGGVECG HGSDSRDEDR VGFYKRYCDI LGVSYGDNLD CGNQSHF |
-Experimental information
Beam | Instrument name: Stanford Synchrotron Radiation Lightsource (SSRL) BL4-2 City: Menlo Park, CA / 国: USA / Type of source: X-ray synchrotron / Wavelength: 1.127 Å / Dist. spec. to detc.: 1.78 mm | ||||||||||||||||||||||||||||||||||||
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Detector | Name: Rayonix MX225-HE | ||||||||||||||||||||||||||||||||||||
Scan | Title: ChiAt1 / Measurement date: Apr 26, 2017 / Storage temperature: 4 °C / Cell temperature: 15 °C / Exposure time: 1 sec. / Unit: 1/A /
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Distance distribution function P(R) | Sofotware P(R): GNOM 5.0 / Number of points: 330 /
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Result | Type of curve: single_conc
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