SULFATASE METABOLISM IN MULTIPLE SULFATASE DEFICIENCY

多种硫酸酯酶缺乏症中的硫酸酯酶代谢

• 批准号: 3315498
• 负责人: ALLEN L HORWITZ
• 金额: $ 14.93万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-12-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3315498
• 关键词: arylsulfatases; fibroblasts; gene expression; genetic mapping; human tissue; hybrid cells; inborn metabolism disorder; lysosomes; messenger RNA; monoclonal antibody; mucopolysaccharidosis type II; sulfatases; tissue /cell culture

The sulfatase deficiency disorders include a number of lysosomal storage diseases and are a significant cause of mental retardation and physical deformities. Since the biochemical and enzymic basis for these disorders have been defined, an important facet of these diseases necessary for diagnostic and therapeutic strategies is the determination of the metabolism of the enzymes involved. Multiple sulfatase deficiency (MSD) is a unique genetic disorder resulting from deficiencies of at least seven distinct sulfatases and appears to result from either defective control of enzymic synthesis or a previously unknown cause of increased degradation of cellular enzymes. The comparison of metabolism of a number of sulfatases in MSD cells with normal or single sulfatase deficiency cells is likely to produce new knowledge of control of levels of cellular hydrolases. The proposed studies will use cultured cells as a source of material. All of the sulfatases will be compared enzymatically to determine what common factors may be necessary for stability of sulfatases, with a view toward defining cellular processes necessary to maintain normal enzyme levels. A major emphasis will be to use antibodies to sulfatases, in particular iduronate sulfatase (IdS-S), to measure rates of synthesis and degradation of this enzyme in cultured cells. If pulse-chase labeling experiments indicate that enzyme degradation is the major defect in MSD, the steps involved in post-translational processing and transport will be investigated. If decreased rates of synthesis are indicated, the factors controlling synthesis, including amounts of translatable mRNA, will be determined. Such probes can also be used for the study of Hunter syndrome (IdS-S deficiency) and its variants and may lead to information about the pathogenesis of mental retardation in this disease. Finally, a genetic approach to the understanding of MSD will be utilized. Established rodent cell lines will be mutagenized and screened for mutants having the MSD phenotype and those which appear to have a genetic defect similar to MSD, as evidenced by lack of complementation in MSD-rodent cell hybrids will be analyzed. Among the uses of such mutants will be the mapping of the human gene responsible for correcting the MSD phenotype to a specific human chromosome. This combination of metabolic studies and genetic analysis is likely to lead to better definition of the controls involved in maintenance of cellular enzyme levels.

硫酸酯酶缺乏症包括许多溶酶体胆积症， 疾病，是智力迟钝和身体发育不良的重要原因。 畸形 因为这些疾病的生化和酶基础 这些疾病的一个重要方面， 诊断和治疗策略是确定 参与酶的代谢。 多发性硫酸酯酶缺乏症（MSD） 一种独特的遗传疾病，由于缺乏至少七种 不同的硫酸酯酶，似乎是由于控制缺陷， 酶合成或以前未知的原因增加降解 细胞酶 几种硫酸酯酶代谢的比较 在具有正常或单一硫酸酯酶缺乏细胞的MSD细胞中， 产生控制细胞水解酶水平的新知识。 拟议的研究将使用培养的细胞作为材料来源。 所有 的硫酸酯酶将进行酶法比较，以确定什么共同的 因素可能是硫酸酯酶稳定性所必需的， 定义维持正常酶水平所必需的细胞过程。 一 主要重点将是使用硫酸酯酶的抗体，特别是 艾杜糖醛酸硫酸酯酶（IdS-S），以测量合成和降解速率 这种酶在培养细胞中的含量。 如果脉冲追踪标记实验 表明酶降解是MSD主要缺陷， 参与翻译后加工和运输的蛋白质将被 研究了 如果显示合成速率降低， 控制合成，包括可翻译mRNA的量， 测定 这种探针也可用于亨特氏综合征的研究 （IdS-S缺陷）及其变体，并可能导致有关 精神发育迟滞的发病机制。 最后，一个基因 将使用了解MSD的方法。 已建立的啮齿动物 将对细胞系进行诱变并筛选具有MSD的突变体 表型和那些似乎具有类似于MSD的遗传缺陷的表型， 如MSD-啮齿动物细胞杂交体中缺乏互补所证明的， 分析了 这些突变体的用途之一将是绘制人类 负责将MSD表型纠正为特定人的基因 染色体 这种代谢研究和遗传分析的结合， 可能导致更好地定义维护中涉及的控制 细胞酶的水平。