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Hypertonic stress modulates eNOS function through O-GlcNAc modification at Thr-866.

Chang Li1,2, An He1, Yongzheng Guo1,2

  • 1Division of Cardiology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016, China.

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|May 29, 2021
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Summary
This summary is machine-generated.

Physical and chemical factors impact endothelial nitric oxide synthase (eNOS) O-GlcNAcylation. Hypertonic stress regulates eNOS O-GlcNAc via the Thr866 site and O-linked N-acetylglucosamine (GlcNAc) transferase (OGT) dependent on AMPK.

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Area of Science:

  • Biochemistry
  • Cellular Biology
  • Physiology

Background:

  • O-GlcNAcylation is an energy-sensitive posttranslational modification regulating endothelial nitric oxide synthase (eNOS) activity.
  • The Thr866 site is crucial for low-glucose-mediated eNOS O-GlcNAc regulation.
  • The role of the Thr866 site in conjunction with other factors influencing eNOS O-GlcNAc remains unclear.

Purpose of the Study:

  • To investigate the effects of various physical and chemical factors on eNOS O-GlcNAcylation and its Thr866 site.
  • To elucidate the underlying mechanism of hypertonic stress-induced eNOS O-GlcNAc modification.

Main Methods:

  • Exposure of cells to hypertonic stress, hyperthermia, hydrogen peroxide, hypoxia, high alcohol concentrations, and low pH.
  • Site-directed mutagenesis of the eNOS Thr866 site.
  • Analysis of eNOS O-GlcNAc levels, O-linked N-acetylglucosamine (GlcNAc) transferase (OGT) expression, and AMP-activated protein kinase (AMPK) activity.

Main Results:

  • Hypertonic stress, hyperthermia, and hydrogen peroxide increased eNOS O-GlcNAc levels; low pH decreased them.
  • The Thr866 site mutation affected eNOS O-GlcNAc levels only under hypertonic conditions.
  • Hypertonic stress upregulated OGT expression via AMPK, which was reversed upon AMPK knockout.

Conclusions:

  • Hypertonic stress, unlike other tested factors, appears to regulate eNOS O-GlcNAc through the Thr866 site.
  • The mechanism involves AMPK-dependent upregulation of OGT expression.
  • This study reveals a novel regulatory pathway for eNOS O-GlcNAcylation under hypertonic stress.