1LDY

HORSE LIVER ALCOHOL DEHYDROGENASE COMPLEXED TO NADH AND CYCLOHEXYL FORMAMIDE (CXF)

1LDY の概要

• エントリーDOI: 10.2210/pdb1ldy/pdb
• 関連するPDBエントリー: 1LDE 3BTO
• 分子名称: ALCOHOL DEHYDROGENASE, ZINC ION, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, ... (5 entities in total)
• 機能のキーワード: dehydrogenase, alcohol, nicotinamide coenzyme, formamides
• 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: 163098.80

構造登録者

Ramaswamy, S.,Plapp, B.V. (登録日: 1996-12-23, 公開日: 1997-04-21, 最終更新日: 2024-02-14)

主引用文献

Ramaswamy, S.,Scholze, M.,Plapp, B.V.
Binding of formamides to liver alcohol dehydrogenase.
Biochemistry, 36:3522-3527, 1997

PubMed Abstract: Amides are analogs of aldehydes and potent inhibitors of liver alcohol dehydrogenases. They can be used for structural studies and for inhibiting the metabolism of alcohols that form toxic products. We studied N-alkyl amides that bind to the enzyme-NADH complex and act as uncompetitive inhibitors against varied concentrations of ethanol (millimolar Kii values, at pH 8 and 25 degrees C): N-propylacetamide (16), delta-valerolactam (1.6), N-formylpiperidine (0.14), N-isobutylformamide (0.028), N-(cyclohexylmethyl)-formamide (0.011), and N-cyclohexylformamide (0.0087). The lower affinity of delta-valerolactam and N-propylacetamide can be explained by steric hindrance with Phe93 of the enzyme. Replacing Phe93 with Ala in the S48T/F93A mutated enzyme, which resembles the natural alpha-isoenzyme of primates, improved binding of delta-valerolactam by 210-fold. The structures of horse liver enzyme complexed with NADH and N-cyclohexylformamide or N-formylpiperidine were determined by X-ray crystallography at 2.5 A resolution. In both complexes, the carbonyl oxygens of the inhibitors bind to the catalytic zinc and form a hydrogen bond to the hydroxyl group of Ser48 of the enzyme. The six-membered rings bind in overlapping, but rotated, positions that optimize hydrophobic interactions. The binding modes of the unreactive formamides appear to resemble the Michaelis complexes of the analogous substrates, with the re face of the carbonyl carbon suitably positioned to accept a hydrogen from NADH.

PubMed: 9132002
DOI: 10.1021/bi962491z

実験手法

X-RAY DIFFRACTION (2.5 Å)