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https://hdl.handle.net/2440/81446
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dc.contributor.author | Tankrathok, A. | - |
dc.contributor.author | Iglesias-Ferna, J. | - |
dc.contributor.author | Luang, S. | - |
dc.contributor.author | Robinson, R. | - |
dc.contributor.author | Kimura, A. | - |
dc.contributor.author | Rovira, C. | - |
dc.contributor.author | Hrmova, M. | - |
dc.contributor.author | Cairns, J. | - |
dc.date.issued | 2013 | - |
dc.identifier.citation | Acta Crystallographica Section D: Biological Crystallography, 2013; 69(10):2124-2135 | - |
dc.identifier.issn | 0907-4449 | - |
dc.identifier.issn | 1399-0047 | - |
dc.identifier.uri | http://hdl.handle.net/2440/81446 | - |
dc.description.abstract | Rice Os7BGlu26 is a GH1 family glycoside hydrolase with a threefold higher kcat/Km value for 4-nitrophenyl β-D-mannoside (4NPMan) compared with 4-nitrophenyl β-D-glucoside (4NPGlc). To investigate its selectivity for β-D-mannoside and β-D-glucoside substrates, the structures of apo Os7BGlu26 at a resolution of 2.20 Å and of Os7BGlu26 with mannose at a resolution of 2.45 Å were elucidated from isomorphous crystals in space group P212121. The (β/α)8-barrel structure is similar to other GH1 family structures, but with a narrower active-site cleft. The Os7BGlu26 structure with D-mannose corresponds to a product complex, with β-D-mannose in the (1)S5 skew-boat conformation. Docking of the (1)S3, (1)S5, (2)SO and (3)S1 pyranose-ring conformations of 4NPMan and 4NPGlc substrates into the active site of Os7BGlu26 indicated that the lowest energies were in the (1)S5 and (1)S3 skew-boat conformations. Comparison of these docked conformers with other rice GH1 structures revealed differences in the residues interacting with the catalytic acid/base between enzymes with and without β-D-mannosidase activity. The mutation of Tyr134 to Trp in Os7BGlu26 resulted in similar kcat/Km values for 4NPMan and 4NPGlc, while mutation of Tyr134 to Phe resulted in a 37-fold higher kcat/Km for 4NPMan than 4NPGlc. Mutation of Cys182 to Thr decreased both the activity and the selectivity for β-D-mannoside. It was concluded that interactions with the catalytic acid/base play a significant role in glycon selection. | - |
dc.description.statementofresponsibility | Anupong Tankrathok, Javier Iglesias-Fernández, Sukanya Luang, Robert C. Robinson, Atsuo Kimura, Carme Rovira, Maria Hrmova and James R. Ketudat Cairns | - |
dc.language.iso | en | - |
dc.publisher | Munksgaard Int Publ Ltd | - |
dc.rights | © 2013 International Union of Crystallography | - |
dc.source.uri | http://dx.doi.org/10.1107/s0907444913020568 | - |
dc.subject | Glycoside Hydrolases | - |
dc.subject | beta-Mannosidase | - |
dc.subject | Crystallography, X-Ray | - |
dc.subject | Mutagenesis, Site-Directed | - |
dc.subject | Catalytic Domain | - |
dc.subject | Protein Conformation | - |
dc.subject | Substrate Specificity | - |
dc.subject | Glycosylation | - |
dc.subject | Hydrolysis | - |
dc.subject | Oryza | - |
dc.title | Structural analysis and insights into the glycon specificity of the rice GH1 Os7BGlu26 β-D-mannosidase | - |
dc.title.alternative | Structural analysis and insights into the glycon specificity of the rice GH1 Os7BGlu26 beta-D-mannosidase | - |
dc.type | Journal article | - |
dc.identifier.doi | 10.1107/S0907444913020568 | - |
pubs.publication-status | Published | - |
dc.identifier.orcid | Hrmova, M. [0000-0002-3545-0605] | - |
Appears in Collections: | Agriculture, Food and Wine publications Aurora harvest |
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