5es3: Difference between revisions
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''' | ==Co-crystal structure of LDH liganded with oxamate== | ||
<StructureSection load='5es3' size='340' side='right' caption='[[5es3]], [[Resolution|resolution]] 2.29Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5es3]] is a 8 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5ES3 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5ES3 FirstGlance]. <br> | |||
</td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=OXM:OXAMIC+ACID'>OXM</scene></td></tr> | |||
<tr id='activity'><td class="sblockLbl"><b>Activity:</b></td><td class="sblockDat"><span class='plainlinks'>[http://en.wikipedia.org/wiki/L-lactate_dehydrogenase L-lactate dehydrogenase], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=1.1.1.27 1.1.1.27] </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=5es3 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5es3 OCA], [http://pdbe.org/5es3 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5es3 RCSB], [http://www.ebi.ac.uk/pdbsum/5es3 PDBsum]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
We developed an efficient, automated 2-step purification protocol for the production of milligram quantities of untagged recombinant rat lactate dehydrogenase A (rLDHA) from E. coli, using the AKTAxpress chromatography system. Cation exchange followed by size exclusion results in average final purity in excess of 93% and yields ~ 14 milligrams per 50 ml of original cell culture in EnPresso B media, in under 8 hrs, including all primary sample processing and column equilibration steps. The protein is highly active and coherent biophysically and a viable alternative to the more problematic human homolog for structural and ligand-binding studies; an apo structure of untagged rLDHA was solved to a resolution 2.29 A (PDB ID 5ES3). Our automated methodology uses generic commercially available pre-packed columns and simple buffers, and represents a robust standard method for the production of milligram amounts of untagged rLDHA, facilitating a novel fragment screening approach for new inhibitors. | |||
A Streamlined, Automated Protocol for the Production of Milligram Quantities of Untagged Recombinant Rat Lactate Dehydrogenase A Using AKTAxpressTM.,Nowicki MW, Blackburn EA, McNae IW, Wear MA PLoS One. 2015 Dec 30;10(12):e0146164. doi: 10.1371/journal.pone.0146164., eCollection 2015. PMID:26717415<ref>PMID:26717415</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5es3" style="background-color:#fffaf0;"></div> | |||
[[Category: | == References == | ||
[[Category: | <references/> | ||
[[Category: | __TOC__ | ||
[[Category: | </StructureSection> | ||
[[Category: Wear, M | [[Category: L-lactate dehydrogenase]] | ||
[[Category: Blackburn, E A]] | |||
[[Category: McNae, I W]] | |||
[[Category: Nowicki, M W]] | |||
[[Category: Wear, M A]] | |||
[[Category: Allostery]] | |||
[[Category: Lactate dehydrogenase]] | |||
[[Category: Oxidoreductase]] | |||
Revision as of 23:11, 13 January 2016
Co-crystal structure of LDH liganded with oxamateCo-crystal structure of LDH liganded with oxamate
Structural highlights
Publication Abstract from PubMedWe developed an efficient, automated 2-step purification protocol for the production of milligram quantities of untagged recombinant rat lactate dehydrogenase A (rLDHA) from E. coli, using the AKTAxpress chromatography system. Cation exchange followed by size exclusion results in average final purity in excess of 93% and yields ~ 14 milligrams per 50 ml of original cell culture in EnPresso B media, in under 8 hrs, including all primary sample processing and column equilibration steps. The protein is highly active and coherent biophysically and a viable alternative to the more problematic human homolog for structural and ligand-binding studies; an apo structure of untagged rLDHA was solved to a resolution 2.29 A (PDB ID 5ES3). Our automated methodology uses generic commercially available pre-packed columns and simple buffers, and represents a robust standard method for the production of milligram amounts of untagged rLDHA, facilitating a novel fragment screening approach for new inhibitors. A Streamlined, Automated Protocol for the Production of Milligram Quantities of Untagged Recombinant Rat Lactate Dehydrogenase A Using AKTAxpressTM.,Nowicki MW, Blackburn EA, McNae IW, Wear MA PLoS One. 2015 Dec 30;10(12):e0146164. doi: 10.1371/journal.pone.0146164., eCollection 2015. PMID:26717415[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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