Melecular genetic study in the pathogenesis of achondroplasia

我们建立了一种快速、重复性好的双向电泳方法，用于筛选遗传性疾病患者成纤维细胞中表达的异常蛋白。在银染色后，对电泳凝胶进行半自动数字化仪-个人计算机分析。它用于检测目前病因不明的遗传性疾病中的异常斑点。在一项初步研究中，一些软骨发育不全患者显示出两个异常点，这些点在其他遗传性个体、健康个体或其他代谢/非代谢性疾病个体中未检测到。然而，经过对大量细胞株的调查，我们发现它们不是软骨发育不全的特异性，而是在一些与这种疾病无关的个体中表达。它们可能代表中性多态性或与特定的身体状况有关。同时，除了对疾病中异常蛋白表达进行筛选外，我们还对遗传性溶酶体疾病的突变基因表达的蛋白进行了分析。每个突变基因在人β-半乳糖苷酶缺乏症中表达不同的突变蛋白质的细胞内周转，并且在人β-半乳糖苷酶缺乏症中观察到明显的表型和基因型之间的关系。

We developed a rapid and reproducible method of two-dimensional electrophoresis for screening of abnormal proteins expressed in fibroblasts from patients with inherited diseases. After silver staining, the electrophoresis gel was subjected to semi-automatic digitizer-personal computer analysis. It was used for detection of abnormal spots in inherited diseases of currently unknown etiology. In a preliminary study, some patients with achondroplasia showed two abnormal spots that were not detected in inherited other individuals, healthy or with other metabolic/nonmetabolic diseases. However, after a survey of a large number of cell strains, we found that they are not specific of achondroplasia but expressed in some individuals not related to this disease. They may represent neutral polymorphism or have connection to specific physical condition. We have not further information about their physiological roles as yet.Simulteneously, in addition to streening of abnormal protein expressio in diseases, we did analysis of proteins expressed by mutant genes responsible for inherited lysosomal diseases. Each mutant gene expressed different intracellular turnover of mutant proteins in human beta-galactosidase deficiency diseases, and a clear relation between phenotype and genotype was observed in human beta-galactosidase deficiency diseases.