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Updated: Jun 21, 2025

In vivo and in vitro Studies of Adaptor-clathrin Interaction
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In vivo and in vitro Studies of Adaptor-clathrin Interaction

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An extended interaction site determines binding between AP180 and AP2 in clathrin mediated endocytosis

Samuel Naudi-Fabra1,2, Carlos A Elena-Real1, Ida Marie Vedel1

  • 1Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle-Straße 10, 13125, Berlin, Germany.

Nature Communications
|July 13, 2024
PubMed

Insights

Researchers elucidated the binding dynamics of AP180, a neuronal protein, with adaptor protein AP2. A key interaction site on AP180 dictates the overall binding, crucial for clathrin-mediated endocytosis.

Area of Science:

  • Molecular Biology
  • Cellular Biology
  • Biochemistry

Background:

  • Clathrin-mediated endocytosis involves complex protein interactions mediated by clathrin-associated sorting proteins (CLASPs).
  • These CLASPs, including neuronal protein AP180, possess intrinsically disordered regions (IDRs) whose functions are not fully understood.
  • AP180's specific role and interaction with other key proteins like AP2 remain partially elusive.

Purpose of the Study:

  • To investigate the interaction between the intrinsically disordered region of neuronal AP180 and the major adaptor protein AP2.
  • To determine the binding dynamics and functional significance of this interaction at atomic resolution.
  • To elucidate the role of AP180-AP2 interactions in the early stages of clathrin-mediated endocytosis.

Main Methods:

  • Nuclear Magnetic Resonance (NMR) spectroscopy was employed to study the interaction between AP180 and AP2.
  • Atomic resolution details of the binding interface and dynamics were analyzed.
  • Binding affinities and the contribution of different interaction sites were characterized.

Main Results:

  • An extended and strong interaction site (approximately 70 residues) within AP180's IDR was identified, significantly determining AP180-AP2 interaction.
  • The interaction exists in a dynamic equilibrium between bound and unbound states.
  • Weaker binding sites contribute to overall affinity at higher AP2 concentrations.

Conclusions:

  • The identified 70-residue interaction site on AP180 plays a central role in recruiting adaptors to clathrin-coated pits.
  • Transient and promiscuous interactions facilitate network reshaping during cargo uptake.
  • Understanding these dynamics provides insights into the precise molecular mechanisms of endocytosis.

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