Abstract
Mounting studies have demonstrated that RAB3GAP1 expression is modified in brain diseases with multiple neurobiological functions and processes and acts as a potentially significant target. However, the cellular and molecular events arising from RAB3GAP1 dysexpression are still incompletely understood. In this work, underexpression and overexpression of RAB3GAP1 were first induced into cultured mouse cortical neurons by transfection with lentivirus plasmids. Then we globally explored the effects of RAB3GAP1 dysexpression on the proteome of the neurons through the use of isobaric tag for relative and absolute quantitation (iTRAQ)-based quantitative proteomics with bioinformatics. A total of 364 proteins in the RAB3GAP1-underexpression group and 314 proteins in the RAB3GAP1-overexpression group were identified to be differentially expressed. Subsequent bioinformatics analysis indicated that the proteome functional expression profiles induced by RAB3GAP1 underexpression and overexpression were different, suggesting the potential differences in biological processes and cellular effects. Subsequent intergroup cross-comparison revealed some candidate target proteins regulated directly by RAB3GAP1. Further parallel reaction monitoring (PRM) analysis illustrated that Sub1, Ssrp1, and Top1 proteins might serve as new potentially important linkers in the RAB3GAP1-mediated autophagy pathway in the cortical neurons. Collectively, the current proteomics data furnished new valuable insights to better understand the regulatory molecular mechanism of neuronal RAB3GAP1.
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The proteomics raw data can be found in the ProteomeXchange Consortium with the identifier PXD023412. The datasets supporting this article have been uploaded as part of the electronic supplementary material.
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This study was supported by the National Natural Science Foundation of China (Grant Nos. 31770890 and 31570826) and the Health and Family Planning Commission of Chongqing Nanan District (Grant No. 2018-03).
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YL, HX, and JZ designed and performed the experiments; YL, FT, DL, and SL performed the experiments; YL, WL, WG, and WC analyzed the data; HX, FY, and JZ supervised and designed the project; YL, RH, and JZ wrote and polished the manuscript. All authors have approved the final version of the manuscript.
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The experimental protocol was approved by the Ethical Committee of Chongqing Medical University. Animals were treated in accordance with the National Institutes of Health Guidelines for the use and care of laboratory animals.
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Liu, Y., Tian, F., Li, S. et al. Global effects of RAB3GAP1 dysexpression on the proteome of mouse cortical neurons. Amino Acids 53, 1339–1350 (2021). https://doi.org/10.1007/s00726-021-03058-9
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DOI: https://doi.org/10.1007/s00726-021-03058-9