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Quantitative Proteomic and Phosphoproteomic Analyses Reveal a Role of Death-Associated Protein Kinase 1 in Regulating Hippocampal Synapse

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Abstract

Death-associated protein kinase 1 (DAPK1) is a stress-responsive calcium/calmodulin (CaM)-regulated serine/threonine protein kinase that is actively involved in stress-induced cell death. The dysregulation of DAPK1 has been established in various neurological disorders such as epilepsy, Alzheimer’s disease (AD), and Parkinson’s disease (PD). Recent research indicates a synaptic localization of DAPK1 in neurons, suggesting a potential role of DAPK1 in modulating synaptic structure and function. However, the key molecules and pathways underlying the influence of DAPK1 on synapses remain elusive. We utilized quantitative proteomic and phosphoproteomic analyses to compare the differences in protein expression and phosphorylation in hippocampal tissues of wild-type (WT) and DAPK1-knockout (KO) mice. Bioinformatic analysis of differentially expressed proteins and phosphoproteins revealed a preferential enrichment of proteins involved in regulating synaptic function, cytoskeletal structure, and neurotransmission. Gene set enrichment analysis (GESA) highlighted altered presynaptic functions including synaptic vesicle priming and glutamate secretion in KO mice. Besides, we observed that proteins with potential phosphorylation motifs of ERK and DAPK1 were overrepresented among the differential phosphoproteins and were highly enriched in neuronal function-related pathways. Furthermore, Western blot analysis validated differences in the expression of several proteins closely associated with presynaptic organization, dendrites and calcium transmembrane transport between KO and WT mice, further corroborating the potential involvement of DAPK1 in the regulation of synaptic functions. Overall, our data provide molecular evidence to elucidate the physiological links between DAPK1 and neuronal functions and help clarify the role of DAPK1 in the pathogenesis of neurodevelopmental and neurodegenerative diseases.

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Data Availability

The original datasets analyzed in the study are included in the supplementary material. Further inquiries are available from the corresponding author upon reasonable request.

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Acknowledgements

We appreciate the animal facility of Fujian Medical University in assisting with animal maintenance.

Funding

The research was supported by the National Natural Science Foundation of China (82001128, 82271449 and 81970993), the Natural Science Foundation of Fujian Province (2021J01672), and the start-up funding of Fujian Medical University (XRCZX2019039).

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  1. Fujian Key Laboratory of Translational Research in Cancer and Neurodegenerative Diseases, School of Basic Medical Sciences, Fujian Medical University, Fuzhou, 350122, Fujian, China

    Yuan Tian, Xiaoqing Zheng, Ruomeng Li, Li Hu, Xindong Shui, Long Wang, Dongmei Chen, Tae Ho Lee & Tao Zhang

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Contributions

THL and TZ conceived this project, designed experiments, analyzed the data, and wrote the manuscript. YT performed the experiments and analyzed the data. XZ, LH, and RL helped with the sample collection. XS, LW, and DC offered technical assistance. All authors have contributed to and approved the final manuscript.

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Correspondence to Tae Ho Lee or Tao Zhang.

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Tian, Y., Zheng, X., Li, R. et al. Quantitative Proteomic and Phosphoproteomic Analyses Reveal a Role of Death-Associated Protein Kinase 1 in Regulating Hippocampal Synapse. Mol Neurobiol 61, 1794–1806 (2024). https://doi.org/10.1007/s12035-023-03674-4

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