CATHECHOL-O-METHYLTRANSFERASE ACTIVITY IN CULTURED HUMAN-SKIN FIBROBLASTS FROM CONTROLS AND PATIENTS WITH DYSTONIA MUSCULORUM DEFORMANS
CATHECHOL-O-METHYLTRANSFERASE ACTIVITY IN CULTURED HUMAN-SKIN FIBROBLASTS FROM CONTROLS AND PATIENTS WITH DYSTONIA MUSCULORUM DEFORMANS
复制标题
DOI：
10.1002/jnr.490060310
复制
发表时间：
1981-01-01
影响因子：
4.2
通讯作者：
GILLER, EL
中科院分区：
文献类型：
--
作者：
BREAKEFIELD, XO;BRAVERMAN, M;GILLER, EL
成纤维细胞提供了一种活细胞来源，其可轻易从人体获取，并用于评估在神经递质和药物代谢中起重要作用的酶的活性的遗传差异。本文描述了培养的人皮肤成纤维细胞匀浆中儿茶酚 - O - 甲基转移酶（COMT，EC 2.1.1.6）活性的生化特征。该酶的许多特性，包括对二羟基苯甲酸和S - 腺苷甲硫氨酸的表观亲和力、最适pH值和（镁离子），以及钙离子的抑制作用，都与人红细胞裂解物中所报道的相似。建立了培养和测定条件，以便对单个成纤维细胞系中的COMT活性进行最佳且可重复的测量。在16个对照细胞系中，COMT活性范围为每毫克蛋白质115 - 263皮摩尔/分钟，平均值为每毫克蛋白质181皮摩尔/分钟。酶活性不随供体的年龄或性别而变化。8例变形性肌张力障碍（一种病因不明的遗传性运动障碍）患者的成纤维细胞中的COMT活性与对照组无显著差异。对12例肌张力障碍患者的细胞系测定了单胺氧化酶（MAO，EC 1.4.3.4）A型活性，其数值与年龄和性别匹配的对照组无显著差异。这两种分解代谢酶活性的遗传变异显然不足以解释该疾病中所描述的单胺代谢的改变。
Fibroblasts provide a source of living cells that can be obtained easily from humans and used to evaluate inherited differences in the activities of enzymes important in neurotransmitter and drug metabolism. Biochemical characteristics of catechol-O-methyltransferase (COMT, EC 2.1.1.6) activity in homogenates of cultured human skin fibroblasts are described. Many properties of the enzyme, including apparent affinity for dihydroxybenzoic acid and S-adenosyl methionine, optimal pH and (Mg2+), and inhibition by Ca2+, are similar to those reported in lysates of human erythrocytes. Culture and assay conditions were established for optimal and reproducible measurement of COMT activity in individual fibroblast lines. In 16 control lines, COMT activity ranged from 115-263 pmol/min per mg protein with a mean of 181 pmol/min per mg protein. Enzyme activity did not vary with the age or sex of the donor. The COMT activities in fibroblasts from 8 patients with dystonia musculorum deformans, an inherited movement disorder of unknown etiology, were not significantly different from controls. Monoamine oxidase (MAO, EC 1.4.3.4) type A activity was measured in 12 lines from patients with dystonia, and values did not differ significantly from age- and sex-matched controls. Inherited variation in activity of these 2 catabolic enzymes apparently is not sufficient to explain alterations in monoamine metabolism described in this disorder.