5m3b: Difference between revisions
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==Structure of cobinamide-bound BtuF mutant W66L, the periplasmic vitamin B12 binding protein in E.coli== | ==Structure of cobinamide-bound BtuF mutant W66L, the periplasmic vitamin B12 binding protein in E.coli== | ||
<StructureSection load='5m3b' size='340' side='right' caption='[[5m3b]], [[Resolution|resolution]] 1.50Å' scene=''> | <StructureSection load='5m3b' size='340' side='right'caption='[[5m3b]], [[Resolution|resolution]] 1.50Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[5m3b]] is a 2 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5M3B OCA]. For a <b>guided tour on the structure components</b> use [ | <table><tr><td colspan='2'>[[5m3b]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5M3B OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5M3B FirstGlance]. <br> | ||
</td></tr><tr id=' | </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=' | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CYN:CYANIDE+ION'>CYN</scene>, <scene name='pdbligand=GOL:GLYCEROL'>GOL</scene></td></tr> | ||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[ | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5m3b FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5m3b OCA], [https://pdbe.org/5m3b PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5m3b RCSB], [https://www.ebi.ac.uk/pdbsum/5m3b PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5m3b ProSAT]</span></td></tr> | ||
</table> | </table> | ||
== Function == | == Function == | ||
[ | [https://www.uniprot.org/uniprot/BTUF_ECOLI BTUF_ECOLI] Part of the ABC transporter complex BtuCDF involved in vitamin B12 import. Binds vitamin B12 and delivers it to the periplasmic surface of BtuC.<ref>PMID:11790740</ref> | ||
<div style="background-color:#fffaf0;"> | <div style="background-color:#fffaf0;"> | ||
== Publication Abstract from PubMed == | == Publication Abstract from PubMed == | ||
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__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: | [[Category: Escherichia coli K-12]] | ||
[[Category: Large Structures]] | |||
[[Category: Korkhov VM]] | |||
[[Category: Kraeutler B]] | |||
[[Category: | [[Category: Locher KP]] | ||
[[Category: | [[Category: Mireku SA]] | ||
[[Category: | [[Category: Ruetz M]] | ||
[[Category: | [[Category: Zhou T]] | ||
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Latest revision as of 12:09, 23 October 2024
Structure of cobinamide-bound BtuF mutant W66L, the periplasmic vitamin B12 binding protein in E.coliStructure of cobinamide-bound BtuF mutant W66L, the periplasmic vitamin B12 binding protein in E.coli
Structural highlights
FunctionBTUF_ECOLI Part of the ABC transporter complex BtuCDF involved in vitamin B12 import. Binds vitamin B12 and delivers it to the periplasmic surface of BtuC.[1] Publication Abstract from PubMedBtuCD-F is an ABC transporter that mediates cobalamin uptake into Escherichia coli. Early in vivo data suggested that BtuCD-F might also be involved in the uptake of cobinamide, a cobalamin precursor. However, neither was it demonstrated that BtuCD-F indeed transports cobinamide, nor was the structural basis of its recognition known. We synthesized radiolabeled cyano-cobinamide and demonstrated BtuCD-catalyzed in vitro transport, which was ATP- and BtuF-dependent. The crystal structure of cobinamide-bound BtuF revealed a conformational change of a tryptophan residue (W66) in the substrate binding cleft compared to the structure of cobalamin-bound BtuF. High-affinity binding of cobinamide was dependent on W66, because mutation to most other amino acids substantially reduced binding. The structures of three BtuF W66 mutants revealed that tight packing against bound cobinamide was only provided by tryptophan and phenylalanine, in line with the observed binding affinities. In vitro transport rates of cobinamide and cobalamin were not influenced by the substitutions of BtuF W66 under the experimental conditions, indicating that W66 has no critical role in the transport reaction. Our data present the molecular basis of the cobinamide versus cobalamin specificity of BtuCD-F and provide tools for in vitro cobinamide transport and binding assays. Conformational Change of a Tryptophan Residue in BtuF Facilitates Binding and Transport of Cobinamide by the Vitamin B12 Transporter BtuCD-F.,Mireku SA, Ruetz M, Zhou T, Korkhov VM, Krautler B, Locher KP Sci Rep. 2017 Jan 27;7:41575. doi: 10.1038/srep41575. PMID:28128319[2] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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