A kinetic assessment of the C-elegans amyloid disaggregation activity enables uncoupling of disassembly and proteolysis
A kinetic assessment of the C-elegans amyloid disaggregation activity enables uncoupling of disassembly and proteolysis
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DOI：
10.1002/pro.234
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发表时间：
2009-11-01
期刊：
影响因子：
8
通讯作者：
Kelly, Jeffery W.
中科院分区：
文献类型：
--
作者：
Bieschke, Jan;Cohen, Ehud;Kelly, Jeffery W.
蛋白质聚集是迟发性神经退行性疾病的共同特征，包括阿尔茨海默病。在阿尔茨海默病中，A β肽的错误组装与与疾病病因相关的蛋白质毒性有遗传联系。A β蛋白毒性的降低部分是由先前报道的所有分解和蛋白水解活性完成的，在热休克因子1的部分控制下，热休克因子1是一种调节细胞质中蛋白质稳态的转录因子，并由胰岛素生长因子信号负调控。在此，我们报告了一种改进的体外测定方法，以定量外源应用秀丽隐杆线虫提取物完成的重组纤维A β分解动力学。通过这个实验，我们证明了秀丽隐杆线虫的A β分解和蛋白水解活性是可分离的。在秀丽隐杆线虫制剂中发现的解聚活性比蛋白水解活性更耐热。在没有蛋白水解的情况下，β分解被发现是一个可逆的过程。未来对分解和蛋白水解活性的分子基础的发现，为延缓衰老性蛋白质毒性提供了希望，这种毒性会导致一系列疾病的神经退行性变。
Protein aggregation is a common feature of late onset neurodegenerative disorders, including Alzheimer's disease. In Alzheimer's disease, misassembly of the A beta peptide is genetically linked to proteotoxicity associated with disease etiology. A reduction in A beta proteotoxicity is accomplished, in part, by the previously reported All disaggregation and proteolysis activities-under partial control of heat shock factor 1, a transcription factor regulating proteostasis in the cytosol and negatively regulated by insulin growth factor signaling. Herein, we report an improved in vitro assay to quantify recombinant fibrillar A beta disaggregation kinetics accomplished by the exogenous application of C. elegans extracts. With this assay we demonstrate that the A beta disaggregation and proteolysis activities of C. elegans are separable. The disaggregation activity found in C. elegans preparations is more heat resistant than the proteolytic activity. A beta disaggregation in the absence of proteolysis was found to be a reversible process. Future discovery of the molecular basis of the disaggregation and proteolysis activities offers the promise of delaying the age-onset proteotoxicity that leads to neurodegeneration in a spectrum of maladies.