Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/118829
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dc.contributor.authorStratilová, B.en
dc.contributor.authorFiráková, Z.en
dc.contributor.authorKlaudiny, J.en
dc.contributor.authorŠesták, S.en
dc.contributor.authorKozmon, S.en
dc.contributor.authorStrouhalová, D.en
dc.contributor.authorGarajová, S.en
dc.contributor.authorAit-Mohand, F.en
dc.contributor.authorHorváthová, Á.en
dc.contributor.authorFarkaš, V.en
dc.contributor.authorStratilová, E.en
dc.contributor.authorHrmova, M.en
dc.date.issued2019en
dc.identifier.citationPlant Molecular Biology, 2019; 100(1-2):1-17en
dc.identifier.issn0167-4412en
dc.identifier.issn1573-5028en
dc.identifier.urihttp://hdl.handle.net/2440/118829-
dc.description.abstractXyloglucan xyloglucosyl transferases (XETs) (EC 2.4.1.207) play a central role in loosening and re-arranging the cellulose-xyloglucan network, which is assumed to be the primary load-bearing structural component of plant cell walls. The full-length sequence of mature TmXET6.3 from Tropaeolum majus (280 residues) was deduced by the nucleotide sequence analysis of near full-length cDNA by Rapid Amplification of cDNA Ends, based on tryptic and chymotryptic peptide sequences. Partly purified TmXET6.3, expressed in Pichia occurred in N-glycosylated and N-deglycosylated forms. The quantification of hetero-transglycosylation activities of TmXET6.3 revealed that (1,3;1,4)-, (1,6)- and (1,4)-β-D-glucooligosaccharides were the preferred acceptor substrates, while (1,4)-β-D-xylooligosaccharides, and arabinoxylo- and glucomanno-oligosaccharides were less preferred. The 3D model of TmXET6.3, and bioinformatics analyses of identified and putative plant xyloglucan endotransglycosylases (XETs)/hydrolases (XEHs) of the GH16 family revealed that H94, A104, Q108, K234 and K237 were the key residues that underpinned the acceptor substrate specificity of TmXET6.3. Compared to the wild-type enzyme, the single Q108R and K237T, and double-K234T/K237T and triple-H94Q/A104D/Q108R variants exhibited enhanced hetero-transglycosylation activities with xyloglucan and (1,4)-β-D-glucooligosaccharides, while those with (1,3;1,4)- and (1,6)-β-D-glucooligosaccharides were suppressed; the incorporation of xyloglucan to (1,4)-β-D-glucooligosaccharides by the H94Q variant was influenced most extensively. Structural and biochemical data of non-specific TmXET6.3 presented here extend the classic XET reaction mechanism by which these enzymes operate in plant cell walls. The evaluations of TmXET6.3 transglycosylation activities and other members of the GH16 family suggested that a broad acceptor substrate specificity in plant XET enzymes could be more widespread than previously anticipated.en
dc.description.statementofresponsibilityBarbora Stratilová, Zuzana Firáková, Jaroslav Klaudiny; Sergej Šesták, Stanislav Kozmon, Dana Strouhalová, Soňa Garajová, Fairouz Ait‑Mohand, Ágnes Horváthová, Vladimír Farkaš, Eva Stratilová, Maria Hrmovaen
dc.language.isoenen
dc.publisherSpringeren
dc.rights© Springer Nature B.V. 2019en
dc.subjectBioinformatics; GH16 family; homo- and hetero-transglycosylation; protein molecular modelling; site-directed mutagenesisen
dc.titleEngineering the acceptor substrate specificity in the xyloglucan endotransglycosylase TmXET6.3 from nasturtium seeds (Tropaeolum majus L.)en
dc.typeJournal articleen
dc.identifier.rmid0030111697en
dc.identifier.doi10.1007/s11103-019-00852-8en
dc.relation.granthttp://purl.org/au-research/grants/arc/DP120100900en
dc.identifier.pubid464821-
pubs.library.collectionAgriculture, Food and Wine publicationsen
pubs.library.teamDS14en
pubs.verification-statusVerifieden
pubs.publication-statusPublisheden
dc.identifier.orcidHrmova, M. [0000-0002-3545-0605]en
Appears in Collections:Agriculture, Food and Wine publications

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