The Ken and Ruth Davee Department of Neurology, Northwestern University Feinberg School of Medicine, Chicago, IL 60611, USA.
STAR Protoc. 2021 Feb 13;2(1):100340. doi: 10.1016/j.xpro.2021.100340. eCollection 2021 Mar 19.
Lysosomes are critical for maintaining protein homeostasis and cellular metabolism. Lysosomal dysfunction and disrupted protein trafficking contribute to cell death in neurodegenerative disorders, including Parkinson's disease and dementia. We describe three complementary protocols-the use of protein glycosylation, western blotting, immunofluorescence, and hydrolase activity measurement-to analyze the trafficking and activity of lysosomal proteins in patient-derived neurons differentiated from iPSCs. These methods should help to identify lysosomal phenotypes in patient-derived cultures and aid the discovery of therapeutics that augment lysosomal function. For complete details on the use and execution of this protocol, please refer to Cuddy et al. (2019).
溶酶体对于维持蛋白质内稳态和细胞代谢至关重要。溶酶体功能障碍和蛋白质运输紊乱导致神经退行性疾病（包括帕金森病和痴呆症）中的细胞死亡。我们描述了三种互补的方案——使用蛋白质糖基化、western blot、免疫荧光和水解酶活性测定来分析源自 iPSC 的患者来源神经元中的溶酶体蛋白的运输和活性。这些方法应该有助于鉴定患者来源培养物中的溶酶体表型，并有助于发现增强溶酶体功能的治疗方法。有关该方案使用和执行的完整详细信息，请参阅 Cuddy 等人。（2019 年）。
