Abstract
Ubiquitin C-terminal hydrolase-L1 (UCH-L1) has been proposed as one of the Parkinson's disease (PD) related genes, but the possible molecular connection between UCH-L1 and PD is not well understood. In this study, we discovered an N-terminal 11 amino acid truncated variant UCH-L1 that we called NT-UCH-L1, in mouse brain tissue as well as in NCI-H157 lung cancer and SH-SY5Y neuroblastoma cell lines. In vivo experiments and hydrogen-deuterium exchange (HDX) with tandem mass spectrometry (MS) studies showed that NT-UCH-L1 is readily aggregated and degraded, and has more flexible structure than UCH-L1. Post-translational modifications including monoubiquitination and disulfide crosslinking regulate the stability and cellular localization of NT-UCH-L1, as confirmed by mutational and proteomic studies. Stable expression of NT-UCH-L1 decreases cellular ROS levels and protects cells from H2O2, rotenone and CCCP-induced cell death. NT-UCH-L1-expressing transgenic mice are less susceptible to degeneration of nigrostriatal dopaminergic neurons seen in the MPTP mouse model of PD, in comparison to control animals. These results suggest that NT-UCH-L1 may have the potential to prevent neural damage in diseases like PD.
Publication types
-
Research Support, Non-U.S. Gov't
MeSH terms
-
Animals
-
Brain / enzymology*
-
Brain / pathology
-
Cell Line, Tumor
-
Deuterium Exchange Measurement
-
Disease Models, Animal
-
Enzyme Stability
-
HeLa Cells
-
Humans
-
Hydrogen Peroxide / pharmacology
-
Male
-
Mice
-
Mice, Inbred C57BL
-
Mice, Transgenic
-
Mitochondria / metabolism*
-
Parkinson Disease / genetics*
-
Parkinson Disease / pathology*
-
Protein Processing, Post-Translational
-
Ubiquitin Thiolesterase / chemistry
-
Ubiquitin Thiolesterase / genetics*
-
Ubiquitin Thiolesterase / metabolism*
Substances
-
Ubiquitin carboxyl-Terminal Hydrolase L-1, human
-
Hydrogen Peroxide
-
Ubiquitin Thiolesterase
Grants and funding
This work was supported by the Global Research Lab Program (No. 2012K1A1A2045441), and the Proteogenomics Research Program (No. 2012M3A9B90036680) of the National Research Foundation of Korea. Kim HJ, Kim HJ, Jeong JE, Kim S, Kim YM, Kim Y, and Seo J were supported by the Brain Korea 21 project. Kim HJ was supported by RP-Grant 2010 of Ewha Womans University. This work was also supported by the Basic Science Research Program through the NRF funded by the Ministry of Education, Science, and Technology (Grant 20090063274) and by the Ministry of Education (No. 2013R1A1A2061412. The funder's had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.