3kof: Difference between revisions
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<StructureSection load='3kof' size='340' side='right'caption='[[3kof]], [[Resolution|resolution]] 1.90Å' scene=''> | <StructureSection load='3kof' size='340' side='right'caption='[[3kof]], [[Resolution|resolution]] 1.90Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>[[3kof]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/ | <table><tr><td colspan='2'>[[3kof]] 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=3KOF OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3KOF 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]] 1.9Å</td></tr> | ||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | |||
<tr id=' | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=3kof FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3kof OCA], [https://pdbe.org/3kof PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3kof RCSB], [https://www.ebi.ac.uk/pdbsum/3kof PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3kof 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=3kof FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3kof OCA], [https://pdbe.org/3kof PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3kof RCSB], [https://www.ebi.ac.uk/pdbsum/3kof PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3kof ProSAT]</span></td></tr> | ||
</table> | </table> | ||
== Function == | == Function == | ||
[https://www.uniprot.org/uniprot/TALB_ECOLI TALB_ECOLI] Transaldolase is important for the balance of metabolites in the pentose-phosphate pathway.[HAMAP-Rule:MF_00492] | |||
== Evolutionary Conservation == | == Evolutionary Conservation == | ||
[[Image:Consurf_key_small.gif|200px|right]] | [[Image:Consurf_key_small.gif|200px|right]] | ||
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==See Also== | ==See Also== | ||
*[[Transaldolase|Transaldolase]] | *[[Transaldolase 3D structures|Transaldolase 3D structures]] | ||
== References == | == References == | ||
<references/> | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: | [[Category: Escherichia coli K-12]] | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Clapes P]] | |||
[[Category: Clapes | [[Category: Gutierrez M]] | ||
[[Category: Gutierrez | [[Category: Samland AK]] | ||
[[Category: Samland | [[Category: Sandalova T]] | ||
[[Category: Sandalova | [[Category: Schneider G]] | ||
[[Category: Schneider | [[Category: Schneider S]] | ||
[[Category: Schneider | [[Category: Sprenger GA]] | ||
[[Category: Sprenger | |||
Latest revision as of 11:19, 6 September 2023
Crystal structure of the double mutant F178Y/R181E of E.coli transaldolase BCrystal structure of the double mutant F178Y/R181E of E.coli transaldolase B
Structural highlights
FunctionTALB_ECOLI Transaldolase is important for the balance of metabolites in the pentose-phosphate pathway.[HAMAP-Rule:MF_00492] Evolutionary Conservation Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf. Publication Abstract from PubMedRecently, we reported on a transaldolase B variant (TalB F178Y) that is able to use dihydroxyacetone (DHA) as donor in aldol reactions. In a second round of protein engineering, we aimed at improving the affinity of this variant towards nonphosphorylated acceptor aldehydes, that is, glyceraldehyde (GA). The anion binding site was identified in the X-ray structure of TalB F178Y where a sulfate ion from the buffer was bound in the active site. Therefore, we performed site-directed saturation mutagenesis at three residues forming the putative phosphate binding site, Arg181, Ser226 and Arg228. The focused libraries were screened for the formation of D-fructose from DHA and d,l-GA by using an adjusted colour assay. The best results with respect to the synthesis of D-fructose were achieved with the TalB F178Y/R181E variant, which exhibited an at least fivefold increase in affinity towards d,l-GA (K(M)=24 mM). We demonstrated that this double mutant can use D-GA, glycolaldehyde and the L-isomer, L-GA, as acceptor substrates. This resulted in preparative synthesis of D-fructose, D-xylulose and L-sorbose when DHA was used as donor. Hence, we engineered a DHA-dependent aldolase that can synthesise the formation of polyhydroxylated compounds from simple and cheap substrates at preparative scale. Redesigning the Active Site of Transaldolase TalB from Escherichia coli: New Variants with Improved Affinity towards Nonphosphorylated Substrates.,Schneider S, Gutierrez M, Sandalova T, Schneider G, Clapes P, Sprenger GA, Samland AK Chembiochem. 2010 Feb 10. PMID:20148428[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. See AlsoReferences
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