Recently, the research team of Li Jia from the Chinese Academy of Sciences Shanghai Institute of Materia Medica and Li Xin from Zhejiang University published their research results on Chemical Science under the title of Deconvoluting nitric oxide – protein interactions with spatially resolved multiple imaging. This study developed a new probe that captures the interaction between gas signaling molecule NO and substrate protein in living cells and has spatiotemporal resolution function, providing new insights for the regulation of low glucose induced neuronal apoptosis by gas signaling molecule NO.
Glucose is a key energy substrate for the brain and plays an important role in maintaining central system function. If the plasma glucose concentration drops below 3.9 mmol/L, it is considered to enter a hypoglycemic state, which will have a significant negative impact on brain function. Recent studies have found that neuronal apoptosis is an important pathological marker of hypoglycemia induced neuronal damage. Activation of the apoptotic pathway further triggers a cascade of hippocampal cell death, which in turn affects cognitive function. However, it is still urgent to investigate which signaling molecules will respond to low glucose and ultimately promote apoptosis.
NO is a highly anticipated gaseous signaling molecule, and the means to explore its involvement in disease processes are still extremely limited. NO is a gas molecule, and conventional protein proximity labeling methods are difficult to study the interaction between gas molecules and proteins. To this end, the team developed a probe NOP-1 that responds to NO and can instantly label proteins to elucidate the role of NO in hypoglycemic induced nerve damage.
The NOP-1 developed in this study is a bifunctional probe capable of real-time imaging of NO in cells and covalently labeling neighboring proteins. This unique design converts instantaneous cellular NO signals into permanent fluorescence staining, which is compatible with immunostaining and multiple imaging, and is superior to the commercial probe DAF-FM DA. The research team used this probe to discover that hypoglycemia induced upregulation of NO promotes nerve damage. In terms of mechanism exploration, probe NOP-1 α- The fluorescence of microtubule protein and tyrosine nitration protein modification overlaps spatially. Meanwhile, the study identified proteins labeled with NOP-1 α- Microtubulin. Through protein pull down experiments and pharmacological manipulation, further research has been conducted to demonstrate that NO undergoes tyrosine nitration α- Microtubulin disrupts the homeostasis of cytoskeletal proteins, ultimately leading to cell apoptosis.
The research work has received support from the National Key R&D Program and the National Natural Science Foundation of China.
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