4A5L

Crystal structure of the thioredoxin reductase from Entamoeba histolytica

4A5L の概要

• エントリーDOI: 10.2210/pdb4a5l/pdb
• 分子名称: THIOREDOXIN REDUCTASE, NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE, SULFATE ION, ... (5 entities in total)
• 機能のキーワード: oxidoreductase, redox metabolism, oxidative stress
• 由来する生物種: ENTAMOEBA HISTOLYTICA
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 70717.20

構造登録者

Podust, L.M. (登録日: 2011-10-25, 公開日: 2012-10-31, 最終更新日: 2024-10-09)

主引用文献

Parsonage, D.,Sheng, F.,Hirata, K.,Debnath, A.,Mckerrow, J.H.,Reed, S.L.,Abagyan, R.,Poole, L.B.,Podust, L.M.
X-Ray Structures of Thioredoxin and Thioredoxin Reductase from Entamoeba Histolytica and Prevailing Hypothesis of the Mechanism of Auranofin Action.
J.Struct.Biol., 194:180-, 2016

PubMed Abstract: The anti-arthritic gold-containing drug Auranofin is lethal to the protozoan intestinal parasite Entamoeba histolytica, the causative agent of human amebiasis, in both culture and animal models of the disease. A putative mechanism of Auranofin action proposes that monovalent gold, Au(I), released from the drug, can bind to the redox-active dithiol group of thioredoxin reductase (TrxR). Au(I) binding in the active site is expected to prevent electron transfer to the downstream substrate thioredoxin (Trx), thus interfering with redox homeostasis in the parasite. To clarify the molecular mechanism of Auranofin action in more detail, we determined a series of atomic resolution X-ray structures for E. histolytica thioredoxin (EhTrx) and thioredoxin reductase (EhTrxR), the latter with and without Auranofin. Only the disulfide-bonded form of the active site dithiol (Cys(140)-Cys(143)) was invariably observed in crystals of EhTrxR in spite of the addition of reductants in various crystallization trials, and no gold was found associated with these cysteines. Non-catalytic Cys(286) was identified as the only site of modification, but further mutagenesis studies using the C286Q mutant demonstrated that this site was not responsible for inhibition of EhTrxR by Auranofin. Interestingly, we obtained both of the catalytically-relevant conformations of this bacterial-like, low molecular weight TrxR in crystals without requiring an engineered disulfide linkage between Cys mutants of TrxR and Trx (as was originally done with Escherichia coli TrxR and Trx). We note that the -CXXC- catalytic motif, even if reduced, would likely not provide space sufficient to bind Au(I) by both cysteines of the dithiol group.

PubMed: 26876147
DOI: 10.1016/J.JSB.2016.02.015

実験手法

X-RAY DIFFRACTION (1.66 Å)