MOLECULAR GENETICS OF LYSOSOMAL DISORDERS

我们接近了突变的特征， 遗传性溶酶体疾病，通过解决遗传和分子 这些酶的合成和细胞内路线的变体。 以戈谢病为模型，我们证明了特异性蛋白质 每个观察到的表型的多态性。 我们进一步证明， 在这些遗传性疾病中观察到的表型异质性可能是一种 不同突变的结果，每种突变都会影响酶的活性， 影响药物的加工、区室化和/或稳定性， 溶酶体水解酶 虽然对具体的理解 负责临床多样性的机制已经增加， 蛋白质分析的结果，许多主要的病理生理 这些方法仍然没有得到很好的描述。 最近，我们扩大了 通过应用重组DNA来理解这些疾病 技术来阐明这些酶的基因结构。 我们分离的cDNA克隆允许更完整的 转录和翻译事件的描述。 这些cDNA 克隆还允许通过以下方法定位该酶的基因： 原位杂交以及基因组克隆的分离。 限制性片段长度多态性（RFLP）已被鉴定。 北方和S1核酸酶分析提供了结构的进一步细节 正常基因和突变基因的区别 分子机制导致 这些疾病中的神经系统参与也已经被 研究了 正常和突变的Western分析和脉冲标记 葡萄糖脑苷脂酶的研究表明，高雪氏症的几种表型 疾病是由不同的突变引起的。 我们还建造了人类 脑文库，其中溶酶体酶的cDNA已被 与世隔绝 将这些基因与非神经组织的基因进行比较 应提供关于组织特异性调节的进一步信息 表情 这项研究的结果将提供一个更合理的 基金会为这些遗传性疾病的治疗努力。

We approached the characterization of the mutations resulting in the inherited lysosomal disorders by addressing the genetic and molecular variants in the synthesis and intracellular routing of these enzymes. Using Gaucher's disease as a model, we demonstrated specific protein polymorphisms for each observed phenotype. We further demonstrated that phenotypic heterogeneity seen within these inherited disorders may be a consequence of different mutations, each affecting enzyme activity and influencing the processing, compartmentalization and/or stability of the lysosomal hydrolases. Although the understanding of the specific mechanisms responsible for clinical diversity has increased as a consequence of protein analyses, many of the primary pathophysiological processes have still not been well described. Recently we extended our understanding of these disorders by the application of recombinant DNA techniques to elucidate the structure of the gene(s) for these enzymes. The cDNA clones that we isolated have permitted the more complete description of the transcriptional and translation events. These cDNA clones have also permitted the localization of the gene for this enzyme by in situ hybridization, as well as the isolation of genomic clones. Restriction fragment length polymorphisms (RFLP's) have been identified. Northern and S1 nuclease analyses provide further details of the structure of the normal and mutant genes. The molecular mechanisms leading to nervous system involvement within these disorders have also been investigated. Western analysis and pulselabelling of normal and mutant glucocerebrosidase demonstrate that the several phenotypes of Gaucher's disease are caused by different mutations. We have also constructed human brain libraries from which cDNA for the lysosomal enzymes has been isolated. A comparison of these genes to those from non-neural tissues should provide further information on the regulation of tissue specific expression. The results of this research will provide a more rational foundation for therapeutic endeavors for these inherited disorders.