Phosphoproteome Analysis Identifies a Synaptotagmin-1-Associated Complex Involved in Ischemic Neuron Injury

Mol Cell Proteomics. 2022 May;21(5):100222. doi: 10.1016/j.mcpro.2022.100222. Epub 2022 Mar 5.

Abstract

Cerebral stroke is one of the leading causes of death in adults worldwide. However, the molecular mechanisms of stroke-induced neuron injury are not fully understood. Here, we obtained phosphoproteomic and proteomic profiles of the acute ischemic hippocampus by LC-MS/MS analysis. Quantitative phosphoproteomic analyses revealed that the dysregulated phosphoproteins were involved in synaptic components and neurotransmission. We further demonstrated that phosphorylation of Synaptotagmin-1 (Syt1) at the Thr112 site in cultured hippocampal neurons aggravated oxygen-glucose deprivation-induced neuronal injury. Immature neurons with low expression of Syt1 exhibit slight neuronal injury in a cerebral ischemia model. Administration of the Tat-Syt1T112A peptide protects neurons against cerebral ischemia-induced injury in vitro and in vivo. Surprisingly, potassium voltage-gated channel subfamily KQT member 2 (Kcnq2) interacted with Syt1 and Annexin A6 (Anxa6) and alleviated Syt1-mediated neuronal injury upon oxygen-glucose deprivation treatment. These results reveal a mechanism underlying neuronal injury and may provide new targets for neuroprotection after acute cerebral ischemia onset.

Keywords: Ca(2+)-dependent membrane-binding protein Anxa6; acute ischemic hippocampus; phosphoproteome and proteome; potassium voltage-gated channel Kcnq2; synaptotagmin-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brain Ischemia* / metabolism
  • Cells, Cultured
  • Chromatography, Liquid
  • Glucose / metabolism
  • Humans
  • Neurons / metabolism
  • Oxygen / metabolism
  • Proteomics*
  • Tandem Mass Spectrometry

Substances

  • Glucose
  • Oxygen