Analysis of lysosomal hydrolase trafficking and activity in human iPSC-derived neuronal models.
Analysis of lysosomal hydrolase trafficking and activity in human iPSC-derived neuronal models.
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DOI：
10.1016/j.xpro.2021.100340
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发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Mazzulli JR
中科院分区：
文献类型：
--
作者：
Cuddy LK;Mazzulli JR
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. Hydrolase maturation can be analyzed by protein glycosylation and western blot Immunofluorescence is used to validate hydrolase trafficking to the lysosome Hydrolase function is assessed in living cells with fluorescent activity probes 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.
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影响因子：
64.8
作者：
Kriks, Sonja;Shim, Jae-Won;Piao, Jinghua;Ganat, Yosif M.;Wakeman, Dustin R.;Xie, Zhong;Carrillo-Reid, Luis;Auyeung, Gordon;Antonacci, Chris;Buch, Amanda;Yang, Lichuan;Beal, M. Flint;Surmeier, D. James;Kordower, Jeffrey H.;Tabar, Viviane;Studer, Lorenz
通讯作者：
Studer, Lorenz
影响因子：
16.2
作者：
Cuddy, Leah K.;Wani, Willayat Y.;Mazzulli, Joseph R.
通讯作者：
Mazzulli, Joseph R.
DOI：
10.1073/pnas.1520335113
发表时间：
2016-02-16
影响因子：
11.1
作者：
Mazzulli, Joseph R.;Zunke, Friederike;Krainc, Dimitri
通讯作者：
Krainc, Dimitri
影响因子：
5.3
作者：
Mazzulli, Joseph R.;Zunke, Friederike;Krainc, Dimitri
通讯作者：
Krainc, Dimitri
影响因子：
158.5
作者：
Kim, J.;Hu, C.;Yu, T. W.
通讯作者：
Yu, T. W.