Structure, Regulation and Mechanism of the Action of Oxidizing Enzyme

黄色素对甲酚甲基羟基酶的细胞色素亚基的初级序列已经确定，黄素蛋白亚基的前52个氨基酸的序列也已经确定。用3A分辨率的X射线结晶学数据对这些数据进行了拟合，并解开了大部分的三级结构。其余的序列正在通过DNA测序来确定。利用闪光光解、停流和稳态动力学方法，已经广泛地研究了黄色素可逆解离为亚单位的过程及其动力学机制。该酶催化的反应序列已被证明包括乙烯基衍生物。MPTP是一种非法度冷丁类似物中的污染物，其神经毒性的生物化学事件已被确定，体内毒性与酶学数据相关。已经发现了一种新的代谢产物载体，它能将MPTP的氧化产物MPP泵入线粒体基质，并对其进行了部分表征。水杨酸羟基酶的动力学研究已经完成，使用一种新的方法来分离该酶。比较了“内”肉碱酰基转移酶和“外”肉碱酰基转移酶的动力学和分子特征，发现过氧化物体可能是外酶的来源。

The primary sequence of the cytochrome subunit of the flavocytochrome p-cresol methylhydroxylase has been determined, as has the sequence of the first 52 amino acids of the flavoprotein subunit. X-ray crystallographic data at 3 A resolution have been fitted to these data and much of the tertiary structure has been unravelled. The rest of the sequence is being determined by DNA sequencing. The reversible dissociation of the flavocytochrome into subunits has been extensively investigated, as has its kinetic mechanism, using flash photolysis, stopped-flow, and steady- state kinetic methods. The reaction sequence catalyzed by the enzyme has been shown to include a vinyl derivative. The biochemical events underlying the neurotoxicity of MPTP, a contaminant in an illicit meperidine analog, has been determined and in vivo toxicity correlated with enzymological data. A new metabolite carrier which pumps MPP+, the oxidation product of MPTP, into the mitochondrial matrix, has been discovered and partially characterized. Extensive kinetic studies have been completed on salicylate hydroxylase, using a novel procedure to isolate the enzyme. The kinetic and molecular characteristics of an "inner" and an "outer" carnitine acyltransferase have been compared and peroxisomes shown to be the likely source of the the outer enzyme.