7qea: Difference between revisions
From Proteopedia
Jump to navigationJump to search
No edit summary |
No edit summary |
||
| (One intermediate revision by the same user not shown) | |||
| Line 4: | Line 4: | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[7qea]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7QEA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7QEA FirstGlance]. <br> | <table><tr><td colspan='2'>[[7qea]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7QEA OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7QEA FirstGlance]. <br> | ||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=B9I:(2~{S},3~{S},4~{S},5~{R},6~{S})-3,4,5-tris(oxidanyl)-6-[(1~{R})-6-oxidanyl-3-oxidanylidene-spiro[2-benzofuran-1,9-xanthene]-3-yl]oxy-oxane-2-carboxylic+acid'>B9I</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</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]] 2.28Å</td></tr> | ||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=ACT:ACETATE+ION'>ACT</scene>, <scene name='pdbligand=B9I:(2~{S},3~{S},4~{S},5~{R},6~{S})-3,4,5-tris(oxidanyl)-6-[(1~{R})-6-oxidanyl-3-oxidanylidene-spiro[2-benzofuran-1,9-xanthene]-3-yl]oxy-oxane-2-carboxylic+acid'>B9I</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</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=7qea FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7qea OCA], [https://pdbe.org/7qea PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7qea RCSB], [https://www.ebi.ac.uk/pdbsum/7qea PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7qea 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=7qea FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7qea OCA], [https://pdbe.org/7qea PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7qea RCSB], [https://www.ebi.ac.uk/pdbsum/7qea PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7qea ProSAT]</span></td></tr> | ||
</table> | </table> | ||
| Line 11: | Line 12: | ||
The abundance of recorded protein sequence data stands in contrast to the small number of experimentally verified functional annotation. Here we screened a million-membered metagenomic library at ultrahigh throughput in microfluidic droplets for beta-glucuronidase activity. We identified SN243, a genuine beta-glucuronidase with little homology to previously studied enzymes of this type, as a glycoside hydrolase 3 family member. This glycoside hydrolase family contains only one recently added beta-glucuronidase, showing that a functional metagenomic approach can shed light on assignments that are currently 'unpredictable' by bioinformatics. Kinetic analyses of SN243 characterized it as a promiscuous catalyst and structural analysis suggests regions of divergence from homologous glycoside hydrolase 3 members creating a wide-open active site. With a screening throughput of >10(7) library members per day, picolitre-volume microfluidic droplets enable functional assignments that complement current enzyme database dictionaries and provide bridgeheads for the annotation of unexplored sequence space. | The abundance of recorded protein sequence data stands in contrast to the small number of experimentally verified functional annotation. Here we screened a million-membered metagenomic library at ultrahigh throughput in microfluidic droplets for beta-glucuronidase activity. We identified SN243, a genuine beta-glucuronidase with little homology to previously studied enzymes of this type, as a glycoside hydrolase 3 family member. This glycoside hydrolase family contains only one recently added beta-glucuronidase, showing that a functional metagenomic approach can shed light on assignments that are currently 'unpredictable' by bioinformatics. Kinetic analyses of SN243 characterized it as a promiscuous catalyst and structural analysis suggests regions of divergence from homologous glycoside hydrolase 3 members creating a wide-open active site. With a screening throughput of >10(7) library members per day, picolitre-volume microfluidic droplets enable functional assignments that complement current enzyme database dictionaries and provide bridgeheads for the annotation of unexplored sequence space. | ||
Functional metagenomic screening identifies an unexpected beta-glucuronidase.,Neun S, Brear P, Campbell E, Tryfona T, El Omari K, Wagner A, Dupree P, Hyvonen M, Hollfelder F Nat Chem Biol. 2022 Oct;18(10):1096-1103. doi: 10.1038/s41589-022-01071-x. Epub, 2022 Jul 7. PMID:35799064<ref>PMID:35799064</ref> | Functional metagenomic screening identifies an unexpected beta-glucuronidase.,Neun S, Brear P, Campbell E, Tryfona T, El Omari K, Wagner A, Dupree P, Hyvonen M, Hollfelder F Nat Chem Biol. 2022 Oct;18(10):1096-1103. doi: 10.1038/s41589-022-01071-x. Epub , 2022 Jul 7. PMID:35799064<ref>PMID:35799064</ref> | ||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
Latest revision as of 06:42, 21 November 2024
Crystal structure of fluorescein-di-Beta-D-glucuronide bound to a mutant of SN243 (D415A)
| ||||||||||||
