PROTEIN DEGRADATION IN INFANTILE NEPHROPATHIC CYSTINOSIS

肾病是一种常染色体隐性遗传病，其特征是 二硫氨基酸半胱氨酸在溶酶体内的积累。我们的 先前的研究表明，从慢性阻塞性肺疾病患者身上培养出的皮肤成纤维细胞 胱氨酸病从两者的降解中积累溶酶体胱氨酸 含有外源和内源二硫化物的蛋白质。我们建议 利用这些成纤维细胞研究二硫化物的还原和降解 以确定内源和外源蛋白质的相对含量 溶酶体参与每类蛋白质的降解。 这些研究需要生产35(S)标记的大鼠血清白蛋白， 和35(S)-胰岛素。它们相互作用的结果是独特的 用十二烷基硫酸钠和十二烷基硫酸钠对囊虫症成纤维细胞标记的蛋白质进行分析 聚丙烯酰胺凝胶电泳法、高效液相色谱法 使用紫外光和放化检测器，以及气相色谱-质谱仪。 该项目应提供与几个基本问题相关的数据 从一个新的角度看待细胞的蛋白质分解。

Nephropathic cystinosis is an autosomal recessive disease characterized by intra-lysosomal accumulation of the disulfide amino acid cystine. Our previous work has shown that skin fibroblasts cultured from patients with cystinosis accumulate lysosomal cystine from the degration of both exogenous and endogenous disulfide containing proteins. We propose to employ these fibroblasts to study the disulfide reduction and degradation of endogenous and exogenous proteins to determine the relative amount of lysosomal participation in the degradation of each class of proteins. These studies will require production of 35(S)-labelled rat serum albumin, and 35(S)-insulin. The results of the interaction of these uniquely labelled proteins with cystinotic fibroblasts will be analyzed both by SDS polyacrylamide gel electrophoresis, high performance liquid chromatography using coupled UV and radio-chemical detectors, and GC-mass spectrometry. This project should provide data relevant to several fundamental questions regarding cellular proteolysis from a novel perspective.