CNS MULTICATALYTIC PROTEINASE COMPLEX

CNS 多催化蛋白酶复合物

• 批准号: 2445784
• 负责人: SHERWIN WILK
• 金额: $ 30.35万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-01 至 1999-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2445784
• 关键词: Baculoviridae; SDS polyacrylamide gel electrophoresis; Sf9 cell line; active sites; central nervous system; chemical conjugate; chemical kinetics; chimeric proteins; chymotrypsin; chymotrypsin inhibitor; endopeptidases; enzyme complex; fluorohydrocarbon; imidazole; ketones; magnesium; molecular cloning; proteasome; protein biosynthesis; protein degradation; protein structure function; recombinant proteins; site directed mutagenesis; transcription factor; ubiquitin; vanadium

The multicatalytic proteinase complex (MPC), also known as the 2OS proteasome, first isolated and characterized in our laboratory fifteen years ago, is a high molecular mass proteolytic molecule found in relatively high concentrations in the cytosol and nucleus of all eukaryotic cells. MPC also serves as the catalytic core of a 26S proteasome which recognizes and degrades ubiquitin-protein conjugates in an ATP-requiring reaction. The explosive growth of research on the proteasomes is accounted for by their unique structural and catalytic properties and by evidence of their essential role in several fundamental cellular processes. Ubiquitin-protein conjugates are a prominent feature in the brains of patients with neurodegenerative diseases. Exposure of a mouse neuronal cell line to a recently synthesized potent inhibitor of the "chymotrypsinlike" activity of MPC leads to accumulation of ubiquitin- protein conjugates. Fundamental questions about the relationship of the structure of MPC to its function and its mechanism of action remain unanswered. Biochemical and molecular biological approaches will be employed to further characterize 2OS and 26S proteasomes in brain. The 26S proteasome will be purified and its catalytic properties defined. Ongoing studies on the expression of the heaviest MPC subunit and its role in protein degradation will be continued. Macromolecular assembly and assignment of specific catalytic functions to individual amino acid residues will be studied by molecular biological techniques. Finally more specific and potent inhibitors will be designed and synthesized. The results of these studies should provide new insights into the mechanism of action and function of a unique macromolecule which plays a significant role in CNS physiology and possibly in CNS pathophysiology.

多催化蛋白酶复合物（MPC），也称为2OS 蛋白酶体，首先在我们的实验室中进行隔离和特征 几年前，是一种高分子质量蛋白水解分子 所有的细胞质和核中相对较高的浓度 真核细胞。 MPC也充当26S的催化核心 蛋白酶体识别并降解泛素 - 蛋白结合物 提取ATP的反应。研究的爆炸性增长 蛋白酶体由其独特的结构和催化来解释 属性和证据表明它们在几个基本中的重要作用 细胞过程。泛素 - 蛋白缀合物是一个突出的特征 在神经退行性疾病患者的大脑中。暴露 小鼠神经元细胞系与最近合成的有效抑制剂 MPC的“类似胰蛋白酶”活性导致泛素的积累 蛋白结合物。关于关系的基本问题 MPC的功能结构及其作用机理仍然存在 未得到答复。生化和分子生物学方法将是 用于进一步表征大脑中的2OS和26S蛋白酶。 26年代 蛋白酶体将被纯化并定义其催化特性。正在进行 研究最重的MPC亚基的表达及其在 蛋白质降解将继续。大分子组件和 将特定催化功能分配给单个氨基酸 残基将通过分子生物学技术研究。最后更多 将设计和合成特定和有效的抑制剂。这 这些研究的结果应提供对机制机制的新见解 独特的大分子的作用和功能，它发挥了重要作用 在CNS生理学以及中枢神经系统病理生理学中的作用。