Nardini M, Ridder I S, Rozeboom H J, Kalk K H, Rink R, Janssen D B, Dijkstra B W
Laboratory of Biophysical Chemistry and BIOSON Research Institute, Department of Chemistry, University of Groningen, Nijenborgh 4, 9747 AG Groningen, The Netherlands.
J Biol Chem. 1999 May 21;274(21):14579-86.
Epoxide hydrolases catalyze the cofactor-independent hydrolysis of reactive and toxic epoxides. They play an essential role in the detoxification of various xenobiotics in higher organisms and in the bacterial degradation of several environmental pollutants. The first x-ray structure of one of these, from Agrobacterium radiobacter AD1, has been determined by isomorphous replacement at 2.1-A resolution. The enzyme shows a two-domain structure with the core having the alpha/beta hydrolase-fold topology. The catalytic residues, Asp107 and His275, are located in a predominantly hydrophobic environment between the two domains. A tunnel connects the back of the active-site cavity with the surface of the enzyme and provides access to the active site for the catalytic water molecule, which in the crystal structure, has been found at hydrogen bond distance to His275. Because of a crystallographic contact, the active site has become accessible for the Gln134 side chain, which occupies a position mimicking a bound substrate. The structure suggests Tyr152/Tyr215 as the residues involved in substrate binding, stabilization of the transition state, and possibly protonation of the epoxide oxygen.
环氧化物水解酶催化活性和有毒环氧化物的非辅因子依赖性水解。它们在高等生物中各种外源化合物的解毒以及几种环境污染物的细菌降解中起着至关重要的作用。其中一种来自放射形土壤杆菌AD1的环氧化物水解酶的首个X射线结构已通过同晶置换法在2.1埃分辨率下确定。该酶呈现出两结构域结构,其核心具有α/β水解酶折叠拓扑结构。催化残基Asp107和His275位于两个结构域之间主要为疏水性的环境中。一条通道将活性位点腔的后部与酶的表面相连,并为催化水分子提供进入活性位点的通道,在晶体结构中,该水分子与His275处于氢键距离。由于晶体学接触,活性位点已可供Gln134侧链进入,该侧链占据了模拟结合底物的位置。该结构表明Tyr152/Tyr215是参与底物结合、过渡态稳定以及可能使环氧化物氧质子化的残基。