5an7: Difference between revisions
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==Structure of the engineered retro-aldolase RA95.5-8F with a bound 1,3- diketone inhibitor== | ==Structure of the engineered retro-aldolase RA95.5-8F with a bound 1,3-diketone inhibitor== | ||
<StructureSection load='5an7' size='340' side='right' caption='[[5an7]], [[Resolution|resolution]] 1.10Å' scene=''> | <StructureSection load='5an7' size='340' side='right' caption='[[5an7]], [[Resolution|resolution]] 1.10Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
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<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=5an7 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5an7 OCA], [http://pdbe.org/5an7 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5an7 RCSB], [http://www.ebi.ac.uk/pdbsum/5an7 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5an7 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=5an7 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5an7 OCA], [http://pdbe.org/5an7 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5an7 RCSB], [http://www.ebi.ac.uk/pdbsum/5an7 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5an7 ProSAT]</span></td></tr> | ||
</table> | </table> | ||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Designing catalysts that achieve the rates and selectivities of natural enzymes is a long-standing goal in protein chemistry. Here, we show that an ultrahigh-throughput droplet-based microfluidic screening platform can be used to improve a previously optimized artificial aldolase by an additional factor of 30 to give a >109 rate enhancement that rivals the efficiency of class I aldolases. The resulting enzyme catalyses a reversible aldol reaction with high stereoselectivity and tolerates a broad range of substrates. Biochemical and structural studies show that catalysis depends on a Lys-Tyr-Asn-Tyr tetrad that emerged adjacent to a computationally designed hydrophobic pocket during directed evolution. This constellation of residues is poised to activate the substrate by Schiff base formation, promote mechanistically important proton transfers and stabilize multiple transition states along a complex reaction coordinate. The emergence of such a sophisticated catalytic centre shows that there is nothing magical about the catalytic activities or mechanisms of naturally occurring enzymes, or the evolutionary process that gave rise to them. | |||
Emergence of a catalytic tetrad during evolution of a highly active artificial aldolase.,Obexer R, Godina A, Garrabou X, Mittl PR, Baker D, Griffiths AD, Hilvert D Nat Chem. 2017 Jan;9(1):50-56. doi: 10.1038/nchem.2596. Epub 2016 Aug 29. PMID:27995916<ref>PMID:27995916</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5an7" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
Revision as of 10:48, 9 March 2017
Structure of the engineered retro-aldolase RA95.5-8F with a bound 1,3-diketone inhibitorStructure of the engineered retro-aldolase RA95.5-8F with a bound 1,3-diketone inhibitor
Structural highlights
Publication Abstract from PubMedDesigning catalysts that achieve the rates and selectivities of natural enzymes is a long-standing goal in protein chemistry. Here, we show that an ultrahigh-throughput droplet-based microfluidic screening platform can be used to improve a previously optimized artificial aldolase by an additional factor of 30 to give a >109 rate enhancement that rivals the efficiency of class I aldolases. The resulting enzyme catalyses a reversible aldol reaction with high stereoselectivity and tolerates a broad range of substrates. Biochemical and structural studies show that catalysis depends on a Lys-Tyr-Asn-Tyr tetrad that emerged adjacent to a computationally designed hydrophobic pocket during directed evolution. This constellation of residues is poised to activate the substrate by Schiff base formation, promote mechanistically important proton transfers and stabilize multiple transition states along a complex reaction coordinate. The emergence of such a sophisticated catalytic centre shows that there is nothing magical about the catalytic activities or mechanisms of naturally occurring enzymes, or the evolutionary process that gave rise to them. Emergence of a catalytic tetrad during evolution of a highly active artificial aldolase.,Obexer R, Godina A, Garrabou X, Mittl PR, Baker D, Griffiths AD, Hilvert D Nat Chem. 2017 Jan;9(1):50-56. doi: 10.1038/nchem.2596. Epub 2016 Aug 29. PMID:27995916[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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