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| <StructureSection load='5zoi' size='340' side='right'caption='[[5zoi]], [[Resolution|resolution]] 3.19Å' scene=''> | | <StructureSection load='5zoi' size='340' side='right'caption='[[5zoi]], [[Resolution|resolution]] 3.19Å' scene=''> |
| == Structural highlights == | | == Structural highlights == |
- | <table><tr><td colspan='2'>[[5zoi]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Atcc_43504 Atcc 43504]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5ZOI OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5ZOI FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5zoi]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Helicobacter_pylori Helicobacter pylori]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5ZOI OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5ZOI FirstGlance]. <br> |
- | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=C06:[[(2S,3R,4S,5R)-5-(2-azanyl-6-oxidanylidene-1H-purin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-oxidanyl-phosphoryl]+[(2S,3S,4S,5R,6R)-6-methyl-3,4,5-tris(oxidanyl)oxan-2-yl]+hydrogen+phosphate'>C06</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]] 3.19Å</td></tr> |
- | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">fucT ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=210 ATCC 43504])</td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=C06:[[(2S,3R,4S,5R)-5-(2-azanyl-6-oxidanylidene-1H-purin-9-yl)-3,4-bis(oxidanyl)oxolan-2-yl]methoxy-oxidanyl-phosphoryl]+[(2S,3S,4S,5R,6R)-6-methyl-3,4,5-tris(oxidanyl)oxan-2-yl]+hydrogen+phosphate'>C06</scene></td></tr> |
- | <tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/4-galactosyl-N-acetylglucosaminide_3-alpha-L-fucosyltransferase 4-galactosyl-N-acetylglucosaminide 3-alpha-L-fucosyltransferase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=2.4.1.152 2.4.1.152] </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=5zoi FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5zoi OCA], [https://pdbe.org/5zoi PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5zoi RCSB], [https://www.ebi.ac.uk/pdbsum/5zoi PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5zoi ProSAT]</span></td></tr> |
- | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5zoi FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5zoi OCA], [http://pdbe.org/5zoi PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5zoi RCSB], [http://www.ebi.ac.uk/pdbsum/5zoi PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5zoi ProSAT]</span></td></tr> | + | |
| </table> | | </table> |
| == Function == | | == Function == |
- | [[http://www.uniprot.org/uniprot/FUCT_HELPX FUCT_HELPX]] Involved in the biosynthesis of the Lewis X (LeX) trisaccharide of the lipopolysaccharide (LPS) O-antigen. Catalyzes the addition of fucose in alpha 1-3 linkage to Gal-beta-1-4-GlcNAc-beta-O-R (LacNAc-R) type II acceptor.<ref>PMID:12676935</ref> <ref>PMID:16150700</ref> <ref>PMID:16800635</ref> <ref>PMID:17251184</ref> <ref>PMID:9261149</ref> | + | [https://www.uniprot.org/uniprot/FUCT_HELPX FUCT_HELPX] Involved in the biosynthesis of the Lewis X (LeX) trisaccharide of the lipopolysaccharide (LPS) O-antigen. Catalyzes the addition of fucose in alpha 1-3 linkage to Gal-beta-1-4-GlcNAc-beta-O-R (LacNAc-R) type II acceptor.<ref>PMID:12676935</ref> <ref>PMID:16150700</ref> <ref>PMID:16800635</ref> <ref>PMID:17251184</ref> <ref>PMID:9261149</ref> |
- | <div style="background-color:#fffaf0;">
| + | |
- | == Publication Abstract from PubMed ==
| + | |
- | Fucosylated glycoconjugates are involved in a variety of physiological and pathological processes. However, economical production of fucosylated drugs and prebiotic supplements has been hampered by the poor catalytic efficiency of fucosyltransferases. Here, we developed a fluorescence-activated cell sorting system that enables the ultrahigh-throughput screening (>10(7) mutants/hour) of such enzymes and designed a companion strategy to assess the screening performance of the system. After three rounds of directed evolution, a mutant M32 of the alpha1,3-FucT from Helicobacter pylori was identified with 6- and 14-fold increases in catalytic efficiency (k cat/K m) for the synthesis of Lewis x and 3'-fucosyllactose, respectively. The structure of the M32 mutant revealed that the S45F mutation generates a clamp-like structure that appears to improve binding of the galactopyranose ring of the acceptor substrate. Moreover, molecular dynamic simulations reveal that helix alpha5, is more mobile in the M32 mutant, possibly explaining its high fucosylation activity.
| + | |
- | | + | |
- | Directed evolution of an alpha1,3-fucosyltransferase using a single-cell ultrahigh-throughput screening method.,Tan Y, Zhang Y, Han Y, Liu H, Chen H, Ma F, Withers SG, Feng Y, Yang G Sci Adv. 2019 Oct 9;5(10):eaaw8451. doi: 10.1126/sciadv.aaw8451. eCollection 2019, Oct. PMID:31633018<ref>PMID:31633018</ref>
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- | | + | |
- | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
| + | |
- | </div>
| + | |
- | <div class="pdbe-citations 5zoi" style="background-color:#fffaf0;"></div>
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| == References == | | == References == |
| <references/> | | <references/> |
| __TOC__ | | __TOC__ |
| </StructureSection> | | </StructureSection> |
- | [[Category: 4-galactosyl-N-acetylglucosaminide 3-alpha-L-fucosyltransferase]] | + | [[Category: Helicobacter pylori]] |
- | [[Category: Atcc 43504]]
| + | |
| [[Category: Large Structures]] | | [[Category: Large Structures]] |
- | [[Category: Tan, Y]] | + | [[Category: Tan Y]] |
- | [[Category: Yang, G]] | + | [[Category: Yang G]] |
- | [[Category: Enzyme]]
| + | |
- | [[Category: Transferase]]
| + | |
| Structural highlights
Function
FUCT_HELPX Involved in the biosynthesis of the Lewis X (LeX) trisaccharide of the lipopolysaccharide (LPS) O-antigen. Catalyzes the addition of fucose in alpha 1-3 linkage to Gal-beta-1-4-GlcNAc-beta-O-R (LacNAc-R) type II acceptor.[1] [2] [3] [4] [5]
References
- ↑ Ma B, Wang G, Palcic MM, Hazes B, Taylor DE. C-terminal amino acids of Helicobacter pylori alpha1,3/4 fucosyltransferases determine type I and type II transfer. J Biol Chem. 2003 Jun 13;278(24):21893-900. doi: 10.1074/jbc.M301704200. Epub, 2003 Apr 3. PMID:12676935 doi:http://dx.doi.org/10.1074/jbc.M301704200
- ↑ Ma B, Lau LH, Palcic MM, Hazes B, Taylor DE. A single aromatic amino acid at the carboxyl terminus of Helicobacter pylori {alpha}1,3/4 fucosyltransferase determines substrate specificity. J Biol Chem. 2005 Nov 4;280(44):36848-56. doi: 10.1074/jbc.M504415200. Epub 2005 , Sep 2. PMID:16150700 doi:http://dx.doi.org/10.1074/jbc.M504415200
- ↑ Lin SW, Yuan TM, Li JR, Lin CH. Carboxyl terminus of Helicobacter pylori alpha1,3-fucosyltransferase determines the structure and stability. Biochemistry. 2006 Jul 4;45(26):8108-16. doi: 10.1021/bi0601297. PMID:16800635 doi:http://dx.doi.org/10.1021/bi0601297
- ↑ Sun HY, Lin SW, Ko TP, Pan JF, Liu CL, Lin CN, Wang AH, Lin CH. Structure and mechanism of Helicobacter pylori fucosyltransferase. A basis for lipopolysaccharide variation and inhibitor design. J Biol Chem. 2007 Mar 30;282(13):9973-82. Epub 2007 Jan 24. PMID:17251184 doi:http://dx.doi.org/10.1074/jbc.M610285200
- ↑ Ge Z, Chan NW, Palcic MM, Taylor DE. Cloning and heterologous expression of an alpha1,3-fucosyltransferase gene from the gastric pathogen Helicobacter pylori. J Biol Chem. 1997 Aug 22;272(34):21357-63. PMID:9261149
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