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| | <StructureSection load='7zf0' size='340' side='right'caption='[[7zf0]], [[Resolution|resolution]] 1.50Å' scene=''> | | <StructureSection load='7zf0' size='340' side='right'caption='[[7zf0]], [[Resolution|resolution]] 1.50Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[7zf0]] is a 1 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7ZF0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7ZF0 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[7zf0]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Sorghum_bicolor Sorghum bicolor]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7ZF0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7ZF0 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=DHR:(2S)-HYDROXY(4-HYDROXYPHENYL)ETHANENITRILE'>DHR</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=UDP:URIDINE-5-DIPHOSPHATE'>UDP</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.5Å</td></tr> |
| - | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[https://en.wikipedia.org/wiki/cyanohydrin_beta-glucosyltransferase cyanohydrin beta-glucosyltransferase], with EC number [https://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.85 2.4.1.85] </span></td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=DHR:(2S)-HYDROXY(4-HYDROXYPHENYL)ETHANENITRILE'>DHR</scene>, <scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=UDP:URIDINE-5-DIPHOSPHATE'>UDP</scene></td></tr> |
| | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=7zf0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7zf0 OCA], [https://pdbe.org/7zf0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7zf0 RCSB], [https://www.ebi.ac.uk/pdbsum/7zf0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7zf0 ProSAT]</span></td></tr> | | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=7zf0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7zf0 OCA], [https://pdbe.org/7zf0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7zf0 RCSB], [https://www.ebi.ac.uk/pdbsum/7zf0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7zf0 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Function == | | == Function == |
| - | [[https://www.uniprot.org/uniprot/HMNGT_SORBI HMNGT_SORBI]] Involved in the biosynthesis of the cyanogenic glucoside dhurrin. Prevents the dissociation and release of toxic hydrogen cyanide. Mandelonitrile, p-hydroxymandelonitrile, benzyl alcohol, benzoic acid and geraniol, but not hydroquinone(1,4-benzenediol), alpha-terpinol, linalool or farnesol are utilized as acceptor substrates. UDP-glucose, but not UDP-xylose or UDP-glucuronic acid can be used as sugar donor.<ref>PMID:10585420</ref> <ref>PMID:11474068</ref> <ref>PMID:15665094</ref>
| + | [https://www.uniprot.org/uniprot/HMNGT_SORBI HMNGT_SORBI] Involved in the biosynthesis of the cyanogenic glucoside dhurrin. Prevents the dissociation and release of toxic hydrogen cyanide. Mandelonitrile, p-hydroxymandelonitrile, benzyl alcohol, benzoic acid and geraniol, but not hydroquinone(1,4-benzenediol), alpha-terpinol, linalool or farnesol are utilized as acceptor substrates. UDP-glucose, but not UDP-xylose or UDP-glucuronic acid can be used as sugar donor.<ref>PMID:10585420</ref> <ref>PMID:11474068</ref> <ref>PMID:15665094</ref> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
| | == Publication Abstract from PubMed == | | == Publication Abstract from PubMed == |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Cyanohydrin beta-glucosyltransferase]] | + | [[Category: Sorghum bicolor]] |
| - | [[Category: Fredslund, F]] | + | [[Category: Fredslund F]] |
| - | [[Category: Putkaradze, N]] | + | [[Category: Putkaradze N]] |
| - | [[Category: Welner, D H]] | + | [[Category: Welner DH]] |
| - | [[Category: Cyanogenic]]
| + | |
| - | [[Category: Dhurrin biosynthesis]]
| + | |
| - | [[Category: Glycosyltransferase]]
| + | |
| - | [[Category: Gt-b fold]]
| + | |
| - | [[Category: Transferase]]
| + | |
| Structural highlights
Function
HMNGT_SORBI Involved in the biosynthesis of the cyanogenic glucoside dhurrin. Prevents the dissociation and release of toxic hydrogen cyanide. Mandelonitrile, p-hydroxymandelonitrile, benzyl alcohol, benzoic acid and geraniol, but not hydroquinone(1,4-benzenediol), alpha-terpinol, linalool or farnesol are utilized as acceptor substrates. UDP-glucose, but not UDP-xylose or UDP-glucuronic acid can be used as sugar donor.[1] [2] [3]
Publication Abstract from PubMed
Cyanogenic glucosides are important defense molecules in plants with useful biological activities in animals. Their last biosynthetic step consists of a glycosylation reaction that confers stability and increases structural diversity and is catalyzed by the UDP-dependent glycosyltransferases (UGTs) of glycosyltransferase family 1. These versatile enzymes have large and varied substrate scopes, and the structure-function relationships controlling scope and specificity remain poorly understood. Here, we report substrate-bound crystal structures and rational engineering of substrate and stereo-specificities of UGT85B1 from Sorghum bicolor involved in biosynthesis of the cyanogenic glucoside dhurrin. Substrate specificity was shifted from the natural substrate (S)-p-hydroxymandelonitrile to (S)-mandelonitrile by combining a mutation to abolish hydrogen bonding to the p-hydroxyl group with a mutation to provide steric hindrance at the p-hydroxyl group binding site (V132A/Q225W). Further, stereo-specificity was shifted from (S) to (R) by substituting four rationally chosen residues within 6 A of the nitrile group (M312T/A313T/H408F/G409A). These activities were compared to two other UGTs involved in the biosynthesis of aromatic cyanogenic glucosides in Prunus dulcis (almond) and Eucalyptus cladocalyx. Together, these studies enabled us to pinpoint factors that drive substrate and stereo-specificities in the cyanogenic glucoside biosynthetic UGTs. The structure-guided engineering of the functional properties of UGT85B1 enhances our understanding of the evolution of UGTs involved in the biosynthesis of cyanogenic glucosides and will enable future engineering efforts towards new biotechnological applications.
Structure-guided engineering of key amino acids in UGT85B1 controlling substrate and stereo-specificity in aromatic cyanogenic glucoside biosynthesis.,Del Giudice R, Putkaradze N, Dos Santos BM, Hansen CC, Crocoll C, Motawia MS, Fredslund F, Laursen T, Welner DH Plant J. 2022 Jul 12. doi: 10.1111/tpj.15904. PMID:35819080[4]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Jones PR, Moller BL, Hoj PB. The UDP-glucose:p-hydroxymandelonitrile-O-glucosyltransferase that catalyzes the last step in synthesis of the cyanogenic glucoside dhurrin in Sorghum bicolor. Isolation, cloning, heterologous expression, and substrate specificity. J Biol Chem. 1999 Dec 10;274(50):35483-91. doi: 10.1074/jbc.274.50.35483. PMID:10585420 doi:http://dx.doi.org/10.1074/jbc.274.50.35483
- ↑ Tattersall DB, Bak S, Jones PR, Olsen CE, Nielsen JK, Hansen ML, Hoj PB, Moller BL. Resistance to an herbivore through engineered cyanogenic glucoside synthesis. Science. 2001 Sep 7;293(5536):1826-8. doi: 10.1126/science.1062249. Epub 2001 Jul, 26. PMID:11474068 doi:http://dx.doi.org/10.1126/science.1062249
- ↑ Kristensen C, Morant M, Olsen CE, Ekstrom CT, Galbraith DW, Moller BL, Bak S. Metabolic engineering of dhurrin in transgenic Arabidopsis plants with marginal inadvertent effects on the metabolome and transcriptome. Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1779-84. doi:, 10.1073/pnas.0409233102. Epub 2005 Jan 21. PMID:15665094 doi:http://dx.doi.org/10.1073/pnas.0409233102
- ↑ Del Giudice R, Putkaradze N, Dos Santos BM, Hansen CC, Crocoll C, Motawia MS, Fredslund F, Laursen T, Welner DH. Structure-guided engineering of key amino acids in UGT85B1 controlling substrate and stereo-specificity in aromatic cyanogenic glucoside biosynthesis. Plant J. 2022 Jul 12. doi: 10.1111/tpj.15904. PMID:35819080 doi:http://dx.doi.org/10.1111/tpj.15904
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