BIOCHEMICAL BASIS OF SULFATASE ACTIVITY

硫酸酯酶活性的生物化学基础

• 批准号: 3324528
• 负责人: ALLEN L HORWITZ
• 金额: $ 3.54万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-05-01 至 1993-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3324528
• 关键词: antibody; arylsulfatases; complementary DNA; electrofocusing; enzyme biosynthesis; enzyme deficiency; enzyme structure; fibroblasts; genetic transcription; genetic translation; human tissue; immunoprecipitation; inborn metabolism disorder; laboratory mouse; laboratory rabbit; laboratory rat; messenger RNA; mucopolysaccharidosis type II; mucopolysaccharidosis type VI; oligosaccharides; protein sequence; regulatory gene; sulfatases; tissue /cell culture; transfection

Mammalian sulfatases are a diverse group of enzymes which hydrolize steroid, mucopolysaccharide and glycolipid sulfates. The sulfatase deficiency disorders include a number of lysosomal storage diseases, and are a significant cause of mental retardation and/or physical deformities. Multiple sulfatase deficiency (MSD) is a unique genetic disorder resulting from deficiencies of at least seven distinct sulfatases and appears to result from increased degradation of cellular sulfatases. These investigations are aimed at the definition of various controls of sulfatase activity and causes of defects at various levels, including transcription, translation and post-translational processes. The elucidation of the defect in MSD is likely to demonstrate new levels of control of cellular hydrolases. Antibody and cDNA probes for steroid sulfatase, iduronate sulfatase (Id-S) and arylsulfatase B (ASB) will be used to study the various levels of control of study the molecular basis of the genetic variation of ASB activity in mouse strains with high or low ASB activity. Synthesis and processing of ASB in these mice using immunoprecipitation and pulse-chase labeling will also be studied to detect the effect of regulatory genes on mouse ASB. cDNA clones and antibodies for human ASB and human Id-S will be isolated and used to characterize the mRNA and newly synthesized polypeptides in ASB deficient Maroteaux-Lamy syndrome and Id-S deficient Hunter disease. These probes plus those for steroid sulfatase will be used to characterize the synthesis and processing of sulfatase in MSD. cDNA for sulfatase will be transfected into MSD and normal cells, and the translation and processing of the transfected STS will be followed. The subcellular compartmentalization and degradation of microinjected STS from normal and MSD cells will be evaluated. Immunoprecipitated labeled sulfatases from normal and MSD cells will be compared by isoelectric focussing, peptide mapping, oligosaccharide content and subcellular location. The role of cellular stabilization factors for sulfatases will be investigated. The route and location of post-translational processing and the role of the Golgi complex in sulfatase processing, including degradation, will be determined. The understanding of all these processes will be important in devising strategies for the treatment of enzyme deficiency diseases.

哺乳动物硫酸酯酶是一组不同的酶， 水解类固醇、粘多糖和硫酸糖脂。 硫酸酯酶缺乏症包括许多溶酶体疾病 贮藏病，是导致智力低下的重要原因 和/或身体畸形。 多种硫酸酯酶缺乏症 (MSD) 是一种独特的遗传性疾病，至少由以下缺陷引起 七种不同的硫酸酯酶，似乎是由于增加 细胞硫酸酯酶的降解。 这些调查的目的是 在硫酸酯酶活性的各种控制的定义和 各种水平缺陷的原因，包括转录， 翻译和翻译后过程。 的阐明 MSD 的缺陷可能会展现出新的控制水平 细胞水解酶。 类固醇硫酸酯酶、艾杜糖醛酸的抗体和 cDNA 探针 硫酸酯酶 (Id-S) 和芳基硫酸酯酶 B (ASB) 将用于研究 不同水平的控制研究的分子基础 高或高小鼠品系中 ASB 活性的遗传变异 ASB 活性低。 这些小鼠体内 ASB 的合成和加工 使用免疫沉淀和脉冲追踪标记也将 研究检测调控基因对小鼠 ASB 的影响。 人 ASB 和人 Id-S 的 cDNA 克隆和抗体将 被分离并用于表征 mRNA 和新 在 ASB 缺陷的 Maroteaux-Lamy 中合成的多肽 综合征和 Id-S 缺陷亨特病。 这些探头加上 类固醇硫酸酯酶的那些将用于表征 MSD 中硫酸酯酶的合成和加工。 硫酸酯酶的cDNA 将被转染到 MSD 和正常细胞中，并进行翻译 随后将处理转染的 STS。 这 亚细胞区室化和降解 将评估来自正常细胞和 MSD 细胞的显微注射 STS。 来自正常细胞和 MSD 细胞的免疫沉淀标记硫酸酯酶 将通过等电聚焦、肽图进行比较， 寡糖含量和亚细胞位置。 的作用 将研究硫酸酯酶的细胞稳定因子。 翻译后处理的路线和位置以及 高尔基复合体在硫酸酯酶加工中的作用，包括 退化，将被确定。 对这一切的理解 流程对于制定战略非常重要 治疗酶缺乏性疾病。

项目成果

Sanfilippo syndrome type D in two adolescent sisters.

两个青春期姐妹的 D 型 Sanfilippo 综合征。

• DOI: 10.1136/jmg.28.6.402
• 发表时间: 1991
• 期刊: Journal of medical genetics
• 影响因子: 4
• 作者: Siciliano,L;Fiumara,A;Pavone,L;Freeman,C;Robertson,D;Morris,CP;Hopwood,JJ;DiNatale,P;Musumeci,S;Horwitz,AL
• 通讯作者: Horwitz,AL

Unusual mucopolysaccharide disorder with corneal and scleral involvement.

不寻常的粘多糖疾病，累及角膜和巩膜。

• DOI: 10.1016/s0002-9394(14)70684-1
• 发表时间: 1990
• 期刊: American journal of ophthalmology
• 影响因子: 4.2
• 作者: Winterbotham,CT;Torczynski,E;Horwitz,AL;Yue,BY;Font,RL
• 通讯作者: Font,RL