Development of a screening system for the pathogenesis of hereditary dominant diseases

利用高分辨率二维电泳、银染色和数字化个人计算机分析，开发了一种快速、可重复的筛选正常和突变人皮肤成纤维细胞蛋白的方法。凝胶上有300多个斑点。等电点的pH值为3.5 ~ 7.5，分子量为10 ~ 150kda。12株正常对照细胞株的247个蛋白点显示出不同程度的定量差异。其中，有12个斑点存在位置或显著的数量变异。然而，在不同年龄、性别或文化条件的群体中，没有明确的变化。支原体感染可在成纤维细胞中诱导6个新的蛋白斑点，其他斑点无变化。该方法对冯氏病、结节性硬化症、软骨发育不全和溶酶体贮积病等遗传病均无定量和定性变化。然而，在3例软骨发育不全（最常见的显性疾病之一）病例中出现了两个新的斑点。另外4例的蛋白模式正常。本研究表明软骨发育不全可能存在遗传异质性，该筛查系统可用于临床实验室遗传显性疾病的研究。

A rapid and reproducible method for screening of normal and mutant human skin fibroblast proteins was developed using high resolutional two-dimensional electrophoresis, silver staining, and digitizer-personal computer analysis. More than 300 spots were visualized on a gel. Iso-electric points were in the range of pH 3.5-7.5, and molecular weights from 10 to 150 kDa.The 247 protein spots chosen for further statistic analysis in 12 normal control cell strains showed quantitative rariations of various degrees. Among them, 12 spots were found to have positional or remarkably quantitative variations. However, no definite variations among groups with different ages, sexes, or culture conditions. Mycoplasma infections was found to induce six new protein spots in fibroblasts without changes of other spots. No quantitative or quali-tative changes were found in several genetic diseases including von Recklinghausen disease, tuberous sclerosis, achondrogenesis and some lysosomal storage diseases by this method. How-ever, two new spots appeared in three cases of achondroplasia, one of the most common dominant diseases. The protein patterns were normal in the other four cases. Our present study revealed that a genetic heterogeneity may be present in achondroplasia and this screening system is useful for the study of hereditary dominant diseases in clinical laboratories.