Guenthner T M, Hammock B D, Vogel U, Oesch F
J Biol Chem. 1981 Apr 10;256(7):3163-6.
Antibodies raised to homogeneous rat liver microsomal epoxide hydrolase were used to distinguish microsomal epoxide hydrolase from epoxide hydrolase of cytosolic origin in mice and rats. Using double diffusion analysis in agarose gels, we show that anti-rat liver microsomal epoxide hydrolase forms a single precipitin line with solubilized microsomes from rat and mouse liver, but no reaction is seen with the corresponding cytosolic fractions. Rat or mouse microsomal epoxide hydrolase activity (using benzo[a]pyrene 4,5-oxide as substrate) can be completely precipitated out of solubilized preparations by the antibody, which is equipotent against rat and mouse microsomal epoxide hydrolase. No precipitation of cytosolic hydrolase activity (using trans-beta-ethyl styrene oxide as substrate) is seen with any concentration of the antibody tested. Thus, in the case of microsomal epoxide hydrolase, extensive immunological cross-reactivity exists between the two species, rat and mouse. In contrast, no cross-reactivity is detectable between cytosolic and microsomal epoxide hydrolase, even when enzymes from the same species are compared. We conclude that microsomal and cytosolic epoxide hydrolase activities represent distinct and immunologically non-cross-reactive protein species.
用针对大鼠肝脏微粒体环氧化物水解酶产生的抗体来区分小鼠和大鼠微粒体来源的环氧化物水解酶与胞质来源的环氧化物水解酶。通过在琼脂糖凝胶中进行双向扩散分析，我们发现抗大鼠肝脏微粒体环氧化物水解酶与大鼠和小鼠肝脏的可溶性微粒体形成单一沉淀线，但与相应的胞质部分无反应。抗体可将大鼠或小鼠微粒体环氧化物水解酶活性（以苯并[a]芘4,5 - 氧化物为底物）从可溶性制剂中完全沉淀出来，该抗体对大鼠和小鼠微粒体环氧化物水解酶具有同等效力。在所测试的任何抗体浓度下，均未观察到胞质水解酶活性（以反式-β-乙基环氧苯为底物）的沉淀。因此，就微粒体环氧化物水解酶而言，大鼠和小鼠这两个物种之间存在广泛的免疫交叉反应性。相比之下，即使比较同一物种的酶，胞质和微粒体环氧化物水解酶之间也未检测到交叉反应性。我们得出结论，微粒体和胞质环氧化物水解酶活性代表不同的、免疫上无交叉反应的蛋白质种类。
