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Deep-Learning-Assisted Single-Molecule Tracking on a Live Cell Membrane.

Qian Wang1, Hua He1, Qian Zhang1

  • 1State Key Laboratory of Heavy Oil Processing and Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, China.

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Summary

Deep learning enhances single-molecule imaging to track protein interactions in real-time. This method precisely distinguishes monomers from complexes, revealing chemokine receptor CXCR4

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Area of Science:

  • Cellular Biology
  • Biophysics
  • Molecular Imaging

Background:

  • Single-molecule fluorescence imaging is vital for studying protein dynamics but struggles with precise identification of interacting proteins on live cell membranes.
  • Heterogeneous protein distribution and interactions complicate real-time tracking of molecular events.

Purpose of the Study:

  • To develop a deep-learning (DL)-assisted single-molecule imaging method for precise, real-time identification and tracking of protein interactions.
  • To investigate the dynamic behavior of chemokine receptor CXCR4 during early signaling stages on live cell membranes.

Main Methods:

  • Developed a DL model utilizing convolutional, max pooling, and fully connected layers for distinguishing protein monomers and complexes.
  • Trained the DL model to achieve >98% accuracy in identifying monomeric versus complexed protein states.
  • Applied the method to track CXCR4 dynamics on live cell membranes upon ligand activation.

Main Results:

  • The DL-assisted imaging method accurately distinguished between monomeric and complexed CXCR4 with high precision.
  • Upon ligand activation, CXCR4 was observed to dynamically form receptor complexes.
  • CXCR4 complexes were internalized 2.5-fold more rapidly than monomers via a clathrin-dependent pathway.

Conclusions:

  • This study presents the first single-molecule level analysis of CXCR4 early signaling on live cell membranes.
  • The developed DL-assisted imaging technique offers a broadly applicable tool for studying protein family dynamics.
  • This method can elucidate the physiological and pathological functions of various proteins.