5ES3

Co-crystal structure of LDH liganded with oxamate

Summary for 5ES3

• Entry DOI: 10.2210/pdb5es3/pdb
• Descriptor: L-lactate dehydrogenase A chain, OXAMIC ACID (3 entities in total)
• Functional Keywords: lactate dehydrogenase, allostery, oxidoreductase
• Biological source: Rattus norvegicus (Rat)
• Cellular location: Cytoplasm: P04642
• Total number of polymer chains: 8
• Total formula weight: 291618.30

Authors

Nowicki, M.W.,Wear, M.A.,McNae, I.W.,Blackburn, E.A. (deposition date: 2015-11-16, release date: 2016-01-13, Last modification date: 2023-09-27)

Primary citation

Nowicki, M.W.,Blackburn, E.A.,McNae, I.W.,Wear, M.A.
A Streamlined, Automated Protocol for the Production of Milligram Quantities of Untagged Recombinant Rat Lactate Dehydrogenase A Using AKTAxpressTM.
Plos One, 10:e0146164-e0146164, 2015

PubMed Abstract: 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 ÄKTAxpress™ 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 Å (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.

PubMed: 26717415
DOI: 10.1371/journal.pone.0146164

Experimental method

X-RAY DIFFRACTION (2.29 Å)