Pinot F, Salaün J P, Bosch H, Lesot A, Mioskowski C, Durst F
CNRS-IBMP, Département d'Enzymologie Cellulaire et Moléculaire, Strasbourg, France.
Biochem Biophys Res Commun. 1992 Apr 15;184(1):183-93. doi: 10.1016/0006-291x(92)91176-q.
A microsomal fraction from etiolated Vicia sativa seedlings incubated aerobically with [1-14C]oleic acid (Z9-octadecenoic acid) or [1-14C]9,10-epoxystearic acid or [1-14C]9,10-dihydroxystearic acid catalyzed the NADPH-dependent formation of hydroxylated metabolites. The chemical structure of compounds formed from oleic, 9,10-epoxystearic or 9,10-dihydroxystearic acids was established by gas chromatography/mass spectra analysis to be 18-hydroxyoleic acid, 18-hydroxy-9,10-epoxystearic acid and 9,10,18-trihydroxystearic acid, respectively. The reactions required O2 and NADPH and were inhibited by carbon monoxide. As expected for monooxygenase reactions involving cytochrome P450, inhibition could be partially reversed by light and all three reactions were inhibited by antibodies raised against NADPH-cytochrome P450 reductase from Jerusalem artichoke. The omega-hydroxylation of the three substrates was enhanced in microsomes from clofibrate induced seedlings.
将黄化的蚕豆幼苗微粒体部分与[1-¹⁴C]油酸(Z9-十八碳烯酸)、[1-¹⁴C]9,10-环氧硬脂酸或[1-¹⁴C]9,10-二羟基硬脂酸进行需氧孵育,可催化依赖NADPH的羟基化代谢产物的形成。通过气相色谱/质谱分析确定,由油酸、9,10-环氧硬脂酸或9,10-二羟基硬脂酸形成的化合物的化学结构分别为18-羟基油酸、18-羟基-9,10-环氧硬脂酸和9,10,18-三羟基硬脂酸。这些反应需要氧气和NADPH,并受到一氧化碳的抑制。正如涉及细胞色素P450的单加氧酶反应所预期的那样,抑制作用可被光部分逆转,并且所有这三个反应都受到针对来自菊芋的NADPH-细胞色素P450还原酶产生的抗体的抑制。在氯贝丁酯诱导的幼苗微粒体中,三种底物的ω-羟基化作用增强。