PROTEIN DEGRADATION IN INFANTILE NEPHROPATHIC CYSTINOSIS

肾病性胱氨酸病是一种常染色体隐性遗传疾病，其特征是： 二硫化物氨基酸胱氨酸的溶酶体内积累。 我们 以前的工作已经表明，从患有糖尿病的患者培养的皮肤成纤维细胞， 胱氨酸病因两者的降解而积累溶酶体胱氨酸 外源性和内源性含二硫键的蛋白质。 我们建议 利用这些成纤维细胞研究二硫键的还原和降解 的内源性和外源性蛋白质，以确定相对量的 溶酶体参与各类蛋白质的降解。 这些研究将需要生产35（S）-标记的大鼠血清白蛋白， 和35（S）-胰岛素。 这些独特的相互作用的结果 将通过SDS和SDS分析具有胱氨酸病成纤维细胞的标记蛋白质。 聚丙烯酰胺凝胶电泳，高效液相色谱 使用耦合的UV和放射化学检测器，和GC-质谱法。 该项目应提供与几个基本问题有关的数据 从一个新的角度来看待细胞蛋白水解。

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.