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| - | [[Image:3ukf.png|left|200px]] | |
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| - | <!-- | + | ==CRYSTAL STRUCTURE OF UDP-galactopyranose mutase from Aspergillus fumigatus in complex with UDPgalp (reduced)== |
| - | The line below this paragraph, containing "STRUCTURE_3ukf", creates the "Structure Box" on the page.
| + | <StructureSection load='3ukf' size='340' side='right'caption='[[3ukf]], [[Resolution|resolution]] 2.50Å' scene=''> |
| - | You may change the PDB parameter (which sets the PDB file loaded into the applet)
| + | == Structural highlights == |
| - | or the SCENE parameter (which sets the initial scene displayed when the page is loaded),
| + | <table><tr><td colspan='2'>[[3ukf]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Aspergillus_fumigatus Aspergillus fumigatus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3UKF OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3UKF FirstGlance]. <br> |
| - | or leave the SCENE parameter empty for the default display.
| + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.5Å</td></tr> |
| - | --> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=FDA:DIHYDROFLAVINE-ADENINE+DINUCLEOTIDE'>FDA</scene>, <scene name='pdbligand=GDU:GALACTOSE-URIDINE-5-DIPHOSPHATE'>GDU</scene>, <scene name='pdbligand=MSE:SELENOMETHIONINE'>MSE</scene></td></tr> |
| - | {{STRUCTURE_3ukf| PDB=3ukf | SCENE= }}
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=3ukf FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3ukf OCA], [https://pdbe.org/3ukf PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3ukf RCSB], [https://www.ebi.ac.uk/pdbsum/3ukf PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3ukf ProSAT]</span></td></tr> |
| | + | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/GLFA_ASPFM GLFA_ASPFM] UDP-galactopyranose mutase, key flavoenzyme of galactofuranose metabolism that catalyzes the 6-to-5 ring contraction of UDP-galactopyranose to UDP-galactofuranose, the donor used by various galacto-furanosyltransferases (PubMed:16207086, PubMed:18552284, PubMed:22334662, PubMed:23036087, PubMed:25412209, PubMed:26836146). Controls the biosynthesis of galactomannan and galactofuranose containing glycoconjugates (PubMed:18552284). The flavin functions as nucleophile, forming a flavin-sugar adduct that facilitates galactose-ring opening and contraction (PubMed:26836146). The binding of UDP-galactopyranose induces profound conformational changes in the enzyme and two loops on opposite sides of the active site move toward each other by over 10 Angstroms to cover the substrate and create a closed active site (PubMed:22334662).<ref>PMID:16207086</ref> <ref>PMID:18552284</ref> <ref>PMID:22334662</ref> <ref>PMID:23036087</ref> <ref>PMID:25412209</ref> <ref>PMID:26836146</ref> |
| | + | <div style="background-color:#fffaf0;"> |
| | + | == Publication Abstract from PubMed == |
| | + | UDP-galactopyranose mutase (UGM) is a flavin-containing enzyme that catalyzes the reversible conversion of UDP-galactopyranose (UDP-Galp) to UDP-galactofuranose (UDP-Galf). As in prokaryotic UGMs, the flavin needs to be reduced for the enzyme to be active. Here we present the first eukaryotic UGM structures from Aspergillus fumigatus (AfUGM). The structures are of UGM alone, with the substrate UDP-Galp and with the inhibitor UDP. Additionally, we report the structures of AfUGM bound to substrate with oxidized and educed flavin. These structures provide insight into substrate recognition and structural changes observed upon substrate binding involving the mobile loops and the critical arginine residues R182 and R327. Comparison with prokaryotic UGM reveals that despite low sequence identity with known prokaryotic UGMs the overall fold is largely conserved. Structural differences between prokaryotic UGM and AfUGM result from inserts in AfUGM. A notable difference from prokaryotic UGMs is that AfUGM contains a third flexible loop (loop III) above the si-face of the isoalloxazine ring that changes position depending on the redox state of the flavin cofactor. This loop flipping has not been observed in prokaryotic UGMs. In addition we have determined the crystals structures and steady-state kinetic constants of the reaction catalyzed by mutants R182K, R327K, R182A and R327A. These results support our hypothesis that R182 and R327 play important roles in stabilizing the diphosphates of the nucleotide sugar and help to facilitate the positioning of the galactose moiety for catalysis. |
| | | | |
| - | ===CRYSTAL STRUCTURE OF UDP-galactopyranose mutase from Aspergillus fumigatus in complex with UDPgalp (reduced)===
| + | Structural insight into the unique substrate binding mechanism and flavin redox state of UDP-galactopyranose mutase from Aspergillus fumigatus.,van Straaten KE, Routier FH, Sanders DA J Biol Chem. 2012 Feb 10. PMID:22334662<ref>PMID:22334662</ref> |
| | | | |
| | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> |
| | + | </div> |
| | + | <div class="pdbe-citations 3ukf" style="background-color:#fffaf0;"></div> |
| | | | |
| - | <!--
| + | ==See Also== |
| - | The line below this paragraph, {{ABSTRACT_PUBMED_22334662}}, adds the Publication Abstract to the page
| + | *[[UDP-galactopyranose mutase 3D structures|UDP-galactopyranose mutase 3D structures]] |
| - | (as it appears on PubMed at http://www.pubmed.gov), where 22334662 is the PubMed ID number.
| + | == References == |
| - | -->
| + | <references/> |
| - | {{ABSTRACT_PUBMED_22334662}}
| + | __TOC__ |
| - | | + | </StructureSection> |
| - | ==About this Structure== | + | |
| - | [[3ukf]] is a 8 chain structure with sequence from [http://en.wikipedia.org/wiki/Aspergillus_fumigatus Aspergillus fumigatus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3UKF OCA]. | + | |
| - | | + | |
| - | ==Reference== | + | |
| - | <ref group="xtra">PMID:022334662</ref><references group="xtra"/> | + | |
| | [[Category: Aspergillus fumigatus]] | | [[Category: Aspergillus fumigatus]] |
| - | [[Category: UDP-galactopyranose mutase]] | + | [[Category: Large Structures]] |
| - | [[Category: Sanders, D A.R.]] | + | [[Category: Sanders DAR]] |
| - | [[Category: Straaten, K E.Van.]] | + | [[Category: Van Straaten KE]] |
| - | [[Category: Fadh2]]
| + | |
| - | [[Category: Flavoenzyme]]
| + | |
| - | [[Category: Isomerase]]
| + | |
| Structural highlights
Function
GLFA_ASPFM UDP-galactopyranose mutase, key flavoenzyme of galactofuranose metabolism that catalyzes the 6-to-5 ring contraction of UDP-galactopyranose to UDP-galactofuranose, the donor used by various galacto-furanosyltransferases (PubMed:16207086, PubMed:18552284, PubMed:22334662, PubMed:23036087, PubMed:25412209, PubMed:26836146). Controls the biosynthesis of galactomannan and galactofuranose containing glycoconjugates (PubMed:18552284). The flavin functions as nucleophile, forming a flavin-sugar adduct that facilitates galactose-ring opening and contraction (PubMed:26836146). The binding of UDP-galactopyranose induces profound conformational changes in the enzyme and two loops on opposite sides of the active site move toward each other by over 10 Angstroms to cover the substrate and create a closed active site (PubMed:22334662).[1] [2] [3] [4] [5] [6]
Publication Abstract from PubMed
UDP-galactopyranose mutase (UGM) is a flavin-containing enzyme that catalyzes the reversible conversion of UDP-galactopyranose (UDP-Galp) to UDP-galactofuranose (UDP-Galf). As in prokaryotic UGMs, the flavin needs to be reduced for the enzyme to be active. Here we present the first eukaryotic UGM structures from Aspergillus fumigatus (AfUGM). The structures are of UGM alone, with the substrate UDP-Galp and with the inhibitor UDP. Additionally, we report the structures of AfUGM bound to substrate with oxidized and educed flavin. These structures provide insight into substrate recognition and structural changes observed upon substrate binding involving the mobile loops and the critical arginine residues R182 and R327. Comparison with prokaryotic UGM reveals that despite low sequence identity with known prokaryotic UGMs the overall fold is largely conserved. Structural differences between prokaryotic UGM and AfUGM result from inserts in AfUGM. A notable difference from prokaryotic UGMs is that AfUGM contains a third flexible loop (loop III) above the si-face of the isoalloxazine ring that changes position depending on the redox state of the flavin cofactor. This loop flipping has not been observed in prokaryotic UGMs. In addition we have determined the crystals structures and steady-state kinetic constants of the reaction catalyzed by mutants R182K, R327K, R182A and R327A. These results support our hypothesis that R182 and R327 play important roles in stabilizing the diphosphates of the nucleotide sugar and help to facilitate the positioning of the galactose moiety for catalysis.
Structural insight into the unique substrate binding mechanism and flavin redox state of UDP-galactopyranose mutase from Aspergillus fumigatus.,van Straaten KE, Routier FH, Sanders DA J Biol Chem. 2012 Feb 10. PMID:22334662[7]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Bakker H, Kleczka B, Gerardy-Schahn R, Routier FH. Identification and partial characterization of two eukaryotic UDP-galactopyranose mutases. Biol Chem. 2005 Jul;386(7):657-61. PMID:16207086 doi:10.1515/BC.2005.076
- ↑ Schmalhorst PS, Krappmann S, Vervecken W, Rohde M, Müller M, Braus GH, Contreras R, Braun A, Bakker H, Routier FH. Contribution of galactofuranose to the virulence of the opportunistic pathogen Aspergillus fumigatus. Eukaryot Cell. 2008 Aug;7(8):1268-77. PMID:18552284 doi:10.1128/EC.00109-08
- ↑ van Straaten KE, Routier FH, Sanders DA. Structural insight into the unique substrate binding mechanism and flavin redox state of UDP-galactopyranose mutase from Aspergillus fumigatus. J Biol Chem. 2012 Feb 10. PMID:22334662 doi:10.1074/jbc.M111.322974
- ↑ Dhatwalia R, Singh H, Solano LM, Oppenheimer M, Robinson RM, Ellerbrock JF, Sobrado P, Tanner JJ. Identification of the NAD(P)H Binding Site of Eukaryotic UDP-Galactopyranose Mutase. J Am Chem Soc. 2012 Oct 4. PMID:23036087 doi:http://dx.doi.org/10.1021/ja308188z
- ↑ Da Fonseca I, Qureshi IA, Mehra-Chaudhary R, Kizjakina K, Tanner JJ, Sobrado P. Contributions of Unique Active Site Residues of Eukaryotic UDP-Galactopyranose Mutases to Substrate Recognition and Active Site Dynamics. Biochemistry. 2014 Dec 2. PMID:25412209 doi:http://dx.doi.org/10.1021/bi501008z
- ↑ Mehra-Chaudhary R, Dai Y, Sobrado P, Tanner JJ. In Crystallo Capture of a Covalent Intermediate in the UDP-Galactopyranose Mutase Reaction. Biochemistry. 2016 Feb 16;55(6):833-6. doi: 10.1021/acs.biochem.6b00035. Epub, 2016 Feb 4. PMID:26836146 doi:http://dx.doi.org/10.1021/acs.biochem.6b00035
- ↑ van Straaten KE, Routier FH, Sanders DA. Structural insight into the unique substrate binding mechanism and flavin redox state of UDP-galactopyranose mutase from Aspergillus fumigatus. J Biol Chem. 2012 Feb 10. PMID:22334662 doi:10.1074/jbc.M111.322974
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