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Identifying the Potential Substrates of the Depalmitoylation Enzyme Acyl-protein Thioesterase 1

[ Vol. 19 , Issue. 5 ]

Author(s):

Huicong Liu, Peipei Yan, Junyan Ren, Can Wu, Wei Yuan, Muding Rao, Zhongjian Zhang* and Eryan Kong*   Pages 364 - 375 ( 12 )

Abstract:


Background: The homeostasis of palmitoylation and depalmitoylation is involved in various cellular processes, the disruption of which induced severe physiological consequences. Acyl-protein thioesterase (APT) and palmitoyl-protein thioesterases (PPT) catalyze the depalmitoylation process. The natural mutation in human PPT1 caused neurodegenerative disease, yet the understanding of APT1 remains to be elucidated. While the deletion of APT1 in mice was embryonic lethal, the decode of its function strictly rely on the identification of its substrates.

Objective: To determine the potential substrates of APT1 by using the generated human APT1 knockout cell line.

Method: The combined techniques of palmitoyl-protein enrichment and mass-spectrometry were used to analysis the different proteins. Palmitoyl-proteins both in HEK293T and APT1-KO cells were extracted by resin-assisted capture (RAC) and data independent acquisition (DIA) quantitative method of proteomics for data collection.

Results: In total, 382 proteins were identified. The gene ontology classification segregated these proteins into diverse biological pathways e.g. endoplasmic reticulum process and ubiquitin mediated proteolysis. A few potential substrates were selected for verification; indeed, major proteins were palmitoylated. Importantly, their levels of palmitoylation were clearly changed in APT1-KO cells. Interestingly, the proliferation of APT1-KO cells escalated dramatically as compared to that of WT cell, which could be rescued by APT1 overexpression.

Conclusion: Our study provided a large scale of potential substrates of APT1 and thus shall facilitate the understanding of its intervened molecular functions.

Keywords:

APT1, palmitoylation, potential substrates, mass spectrometry, data-independent acquisition, proliferation.

Affiliation:

Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang, Institute of Psychiatry and Neuroscience, Xinxiang Medical University, Xinxiang



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