Establishment of a method of analyzing ATP7B function.

威尔逊病是一种铜代谢的遗传性疾病，由铜转运P型atp酶（ATP7B）缺陷引起。基因分析已被用于肝豆状核变性的诊断。分析ATP7B的功能也将有助于该病的诊断。然而，ATP7B功能的分析方法尚未见报道。我们试图建立一种分析ATP7B功能的方法。首先，我们研究了由另一种铜转运atp酶（ATP7A）引起的Menkes病患者的培养成纤维细胞对ATP7A功能的影响。【材料与方法】培养Menkes病患者、Wilson病患者及对照组成纤维细胞。将收获的细胞匀浆并离心。使用上清液。样品中加入谷胱甘肽- cu溶液、ATP溶液，进行混合。然后，将混合物加入荧光素酶试剂中，然后使用Auto Limat测量混合物中ATP的浓度。【结果与讨论】对照细胞中ATP7A/B的活性为10 ~ 200pmol/mg.pro。/min，表示正常控制的液位大宽。在门克斯病患者的细胞中，活性水平也很大；部分患者的活动水平低于正常范围，但部分患者的活动水平在正常范围内。我们得出结论，这种方法不能用于测量ATP7A/B功能的活性。

Wilson disease is a genetic disorder of copper metabolism caused by a defect in a copper-transporting P type ATPase (ATP7B). Gene analysis has been used for the diagnosis of Wilson disease. Analysis of ATP7B function will also be useful for diagnosis of this disease. However, no method for analyzing of ATP7B function has been reported. We investigated to establish a method for analyzing of ATP7B function. At first we studied about ATP7A function with cultured fibroblasts from patients with Menkes disease that is caused by another copper-transporting ATPase (ATP7A).【Materials and Methods】 Cultured fibroblasts from patients with Menkes disease, patients with Wilson disease and control. Harvested cells were homogenized and centrifuged. The supernatants were used. A glutathione-Cu solution, ATP solution were added to the samples, and mixed. After then, the mixture was added to Luciferase reagent, and then the amount of ATP concentration in the mixture was measured using an Auto Limat.【Results and Discussion】The activity of ATP7A/B in the control cells was 10-200pmol/mg.pro./min, showing that the level of normal control is large wide. In the cells from patients with Menkes disease, the activity levels were also large wide ; in some patients, the activity was lower than the normal ranges, but in some of them, the level was within the normal range. We concluded that this method is not available to measure the activity of ATP7A/B function.