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Development of a Genetically Encoded Sensor for Arginine.

Chun Wang1,2,3,4, Xiaoxue Zhang3,4, Haoyu Mao3,4,5

  • 1School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China.

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|January 21, 2025
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Summary

A new sensor, ArgS1, enables real-time measurement of l-arginine (Arg) in cells. This tool revealed that Arg levels impact breast cancer cell viability, offering insights for Arg depletion therapy.

Keywords:
biosensorcell imagingfluorescent probefluorescent proteinl-arginine

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Medical Diagnostics

Background:

  • L-arginine (Arg) is crucial for metabolic and physiological functions, with altered levels linked to diseases.
  • Current Arg sensors lack specificity, often cross-reacting with l-ornithine or l-lysine.
  • Accurate, real-time Arg measurement is vital for understanding its role in health and disease.

Purpose of the Study:

  • To develop and validate a novel sensor, ArgS1, for specific and real-time detection of l-arginine.
  • To investigate the role of Arg in MDA-MB-231 breast cancer cells using the ArgS1 sensor.
  • To correlate Arg levels with cell viability in the context of arginino-succinate synthetase 1 (ASS1) expression.

Main Methods:

  • Development of ArgS1, a sensor exhibiting concentration-dependent fluorescence changes for Arg.
  • Characterization of ArgS1's affinity (∼64 μM) and dynamic range (ΔR/R0 of 3).
  • Application of ArgS1 to monitor intracellular Arg levels in MDA-MB-231 cells under varying Arg availability and ASS1 expression.

Main Results:

  • ArgS1 demonstrated specific and sensitive detection of Arg in both cytoplasm and organelles.
  • Arg levels in MDA-MB-231 cells decreased upon extracellular Arg depletion, correlating with reduced cell viability.
  • Overexpression of ASS1 led to increased Arg levels and enhanced cell viability.

Conclusions:

  • ArgS1 is an effective tool for real-time monitoring of Arg in human cells across physiological and pathological ranges.
  • Arg levels significantly influence MDA-MB-231 breast cancer cell viability.
  • ArgS1 provides valuable insights for developing Arg-targeted cancer therapies.