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| | <StructureSection load='5es3' size='340' side='right'caption='[[5es3]], [[Resolution|resolution]] 2.29Å' scene=''> | | <StructureSection load='5es3' size='340' side='right'caption='[[5es3]], [[Resolution|resolution]] 2.29Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5es3]] is a 8 chain structure with sequence from [http://en.wikipedia.org/wiki/Buffalo_rat Buffalo rat]. 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://proteopedia.org/fgij/fg.htm?mol=5ES3 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5es3]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Rattus_norvegicus Rattus norvegicus]. 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 [https://proteopedia.org/fgij/fg.htm?mol=5ES3 FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=OXM:OXAMIC+ACID'>OXM</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]] 2.29Å</td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">Ldha, Ldh-1, Ldh1 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=10116 Buffalo rat])</td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><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'>[https://proteopedia.org/fgij/fg.htm?mol=5es3 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5es3 OCA], [https://pdbe.org/5es3 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5es3 RCSB], [https://www.ebi.ac.uk/pdbsum/5es3 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5es3 ProSAT]</span></td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/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], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5es3 ProSAT]</span></td></tr> | + | |
| | </table> | | </table> |
| | + | == Function == |
| | + | [https://www.uniprot.org/uniprot/LDHA_RAT LDHA_RAT] |
| | <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: Buffalo rat]] | |
| - | [[Category: L-lactate dehydrogenase]] | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Blackburn, E A]] | + | [[Category: Rattus norvegicus]] |
| - | [[Category: McNae, I W]] | + | [[Category: Blackburn EA]] |
| - | [[Category: Nowicki, M W]] | + | [[Category: McNae IW]] |
| - | [[Category: Wear, M A]] | + | [[Category: Nowicki MW]] |
| - | [[Category: Allostery]] | + | [[Category: Wear MA]] |
| - | [[Category: Lactate dehydrogenase]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| Structural highlights
Function
LDHA_RAT
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[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
See Also
References
- ↑ Nowicki MW, Blackburn EA, McNae IW, Wear MA. A Streamlined, Automated Protocol for the Production of Milligram Quantities of Untagged Recombinant Rat Lactate Dehydrogenase A Using AKTAxpressTM. PLoS One. 2015 Dec 30;10(12):e0146164. doi: 10.1371/journal.pone.0146164., eCollection 2015. PMID:26717415 doi:http://dx.doi.org/10.1371/journal.pone.0146164
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