5ES3

Co-crystal structure of LDH liganded with oxamate

5ES3 の概要

• エントリーDOI: 10.2210/pdb5es3/pdb
• 分子名称: L-lactate dehydrogenase A chain, OXAMIC ACID (3 entities in total)
• 機能のキーワード: lactate dehydrogenase, allostery, oxidoreductase
• 由来する生物種: Rattus norvegicus (Rat)
• 細胞内の位置: Cytoplasm: P04642
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 291618.30

構造登録者

Nowicki, M.W.,Wear, M.A.,McNae, I.W.,Blackburn, E.A. (登録日: 2015-11-16, 公開日: 2016-01-13, 最終更新日: 2023-09-27)

主引用文献

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

実験手法

X-RAY DIFFRACTION (2.29 Å)