1BTO

HORSE LIVER ALCOHOL DEHYDROGENASE COMPLEXED TO NADH AND (1S,3R)3-BUTYLTHIOLANE 1-OXIDE

1BTO の概要

• エントリーDOI: 10.2210/pdb1bto/pdb
• 分子名称: LIVER ALCOHOL DEHYDROGENASE, ZINC ION, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, ... (5 entities in total)
• 機能のキーワード: oxidoreductase, alcohol, nicotinamide coenzyme, sulfoxide
• 由来する生物種: Equus caballus (horse)
• 細胞内の位置: Cytoplasm: P00327
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 163231.17

構造登録者

Ramaswamy, S.,Plapp, B.V. (登録日: 1996-11-08, 公開日: 1997-04-01, 最終更新日: 2024-05-22)

主引用文献

Cho, H.,Ramaswamy, S.,Plapp, B.V.
Flexibility of liver alcohol dehydrogenase in stereoselective binding of 3-butylthiolane 1-oxides.
Biochemistry, 36:382-389, 1997

PubMed Abstract: Thiolane 1-oxides are analogs of the carbonyl substrates that bind to the alcohol dehydrogenase-NADH complex and are potent uncompetitive inhibitors against alcohol [Chadha, V. K., et al. (1985) J. Med. Chem. 28, 36-40]. The four stereoisomers of 3-butylthiolane 1-oxide (BTO) were separated by chiral phase chromatography. CD and 1H-NMR spectra identified the enantiomeric pairs. 1H-NMR chemical shifts were assigned on the basis of COSY spectra of both diastereoisomers and confirmed by HMQC spectra. Coupling constants were determined through one-dimensional decoupling experiments. NMR with chiral shift reagents, Eu(hfc)3 [europium tris [3-[(heptafluoropropyl)hydroxymethylene]-(+)-camphorate]] or (R)-(-)-N-(3,5-dinitrobenzoyl)-alpha-methylbenzylamine, determined that the most inhibitory isomer is either 1S,3R or 1R,3S. The chemical shifts of protons in the thiolane 1-oxide ring were influenced by the whole structure and were not correlated with the computed Mulliken charges. X-ray crystallography at 2.1 and 1.66 A resolution of the ternary enzyme complexes with NADH demonstrated that the absolute configuration of the most inhibitory (Kii = 0.31 microM) stereoisomer is 1S,3R and the next best inhibitor (Kii = 0.73 microM) is 1S,3S. The thiolane 1-oxide rings bind in the same position, in the substrate binding site, but the geometry of the complexes suggests that the sulfoxides are not transition state analogs. Significantly, the butyl groups of the two isomers are accommodated differently by flexible amino acid side chains adopting alternative rotameric conformations.

PubMed: 9003191
DOI: 10.1021/bi9624604

実験手法

X-RAY DIFFRACTION (2 Å)