Huicong Liu, Peipei Yan, Junyan Ren, Can Wu, Wei Yuan, Muding Rao, Zhongjian Zhang* and Eryan Kong* Pages 364 - 375 ( 12 )
<P>Background: The homeostasis of palmitoylation and depalmitoylation is involved in various cellular processes, the disruption of which induces 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 turned out to be potentially embryonically lethal, the decoding of its function strictly relied on the identification of its substrates. <P> Objective: To determine the potential substrates of APT1 by using the generated human APT1 knockout cell line. <P> Methods : The combined techniques of palmitoyl-protein enrichment and massspectrometry were used to analyze 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. <P> 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 the WT cells, which could be rescued by APT1 overexpression. <P> Conclusion: Our study provides a large scale of potential substrates of APT1, thus facilitating the understanding of its intervened molecular functions.</P>
APT1, palmitoylation, potential substrates, mass spectrometry, data-independent acquisition, proliferation.
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