1E3E

Mouse class II alcohol dehydrogenase complex with NADH

1E3E の概要

• エントリーDOI: 10.2210/pdb1e3e/pdb
• 分子名称: ALCOHOL DEHYDROGENASE, CLASS II, 1,4-DIHYDRONICOTINAMIDE ADENINE DINUCLEOTIDE, ZINC ION, ... (4 entities in total)
• 機能のキーワード: oxidoreductase
• 由来する生物種: MUS MUSCULUS (HOUSE MOUSE)
• 細胞内の位置: Cytoplasm : Q9QYY9
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 81781.35

構造登録者

Svensson, S.,Hoeoeg, J.O.,Schneider, G.,Sandalova, T. (登録日: 2000-06-14, 公開日: 2000-09-12, 最終更新日: 2023-12-13)

主引用文献

Svensson, S.,Hoeoeg, J.O.,Schneider, G.,Sandalova, T.
Crystal Structure of Mouse Class II Alcohol Dehydrogenase Reveal Determinants of Substrate Specificity and Catalytic Efficiency
J.Mol.Biol., 302:441-, 2000

PubMed Abstract: The structure of mouse class II alcohol dehydrogenase (ADH2) has been determined in a binary complex with the coenzyme NADH and in a ternary complex with both NADH and the inhibitor N-cyclohexylformamide to 2.2 A and 2.1 A resolution, respectively. The ADH2 dimer is asymmetric in the crystal with different orientations of the catalytic domains relative to the coenzyme-binding domains in the two subunits, resulting in a slightly different closure of the active-site cleft. Both conformations are about half way between the open apo structure and the closed holo structure of horse ADH1, thus resembling that of ADH3. The semi-open conformation and structural differences around the active-site cleft contribute to a substantially different substrate-binding pocket architecture as compared to other classes of alcohol dehydrogenase, and provide the structural basis for recognition and selectivity of alcohols and quinones. The active-site cleft is more voluminous than that of ADH1 but not as open and funnel-shaped as that of ADH3. The loop with residues 296-301 from the coenzyme-binding domain is short, thus opening up the pocket towards the coenzyme. On the opposite side, the loop with residues 114-121 stretches out over the inter-domain cleft. A cavity is formed below this loop and adds an appendix to the substrate-binding pocket. Asp301 is positioned at the entrance of the pocket and may control the binding of omega-hydroxy fatty acids, which act as inhibitors rather than substrates. Mouse ADH2 is known as an inefficient ADH with a slow hydrogen-transfer step. By replacing Pro47 with His, the alcohol dehydrogenase activity is restored. Here, the structure of this P47H mutant was determined in complex with NADH to 2.5 A resolution. His47 is suitably positioned to act as a catalytic base in the deprotonation of the substrate. Moreover, in the more closed subunit, the coenzyme is allowed a position closer to the catalytic zinc. This is consistent with hydrogen transfer from an alcoholate intermediate where the Pro/His replacement focuses on the function of the enzyme.

PubMed: 10970744
DOI: 10.1006/JMBI.2000.4039

実験手法

X-RAY DIFFRACTION (2.12 Å)