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Related Experiment Video

Updated: Apr 18, 2026

Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
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PROTAC internalization and target degradation require clathrin-mediated endocytosis.

Hao-Yang Liu1, Zhengyu Wang2, Rahul Sharma1

  • 1Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX, United States.

Biorxiv : the Preprint Server for Biology
|April 17, 2026
PubMed
Summary

Proteolysis-targeting chimeras (PROTACs) enter cells via clathrin-mediated endocytosis, not passive diffusion. This pathway is essential for PROTAC function and targeted protein degradation.

Keywords:
CD36Proteolysis Targeting Chimerascellular uptakeclathrin-mediated endocytosisprotein degradation

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Area of Science:

  • Cellular Biology
  • Molecular Pharmacology
  • Drug Delivery

Background:

  • Proteolysis-targeting chimeras (PROTACs) are novel therapeutics for targeted protein degradation.
  • The cellular entry mechanism for PROTACs is currently unknown.
  • CD36 has been identified as a PROTAC receptor, but its uptake pathway is unclear.

Purpose of the Study:

  • To elucidate the cellular uptake mechanism of PROTACs.
  • To determine the role of CD36 in PROTAC internalization.
  • To investigate the pathway governing PROTAC-induced protein degradation.

Main Methods:

  • Live-cell imaging to visualize PROTAC and CD36 trafficking.
  • Identification of CD36 C-terminal motifs.
  • Genetic and pharmacological disruption of clathrin-mediated endocytosis.

Main Results:

  • PROTAC uptake and function depend on clathrin-mediated endocytosis.
  • Novel clathrin adaptor-binding motifs were identified in CD36.
  • CD36 and PROTACs colocalize at clathrin-coated pits.
  • Disruption of clathrin assembly abrogates PROTAC-mediated protein degradation.

Conclusions:

  • Clathrin-mediated endocytosis is the primary mechanism for PROTAC cellular entry.
  • This pathway is critical for the efficacy of diverse PROTACs.
  • Understanding this mechanism can optimize targeted degrader design and cellular response.