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AP-2/Eps15 interaction is required for receptor-mediated endocytosis
A Benmerah1, C Lamaze, B Bègue
1Developpement Normal et Pathologique du Systeme Immunitaire, Institut National de la Sante et de la Recherche Medicale U 429, Hopital Necker-Enfants Malades, 75743 Paris Cedex 15, France.
The Journal of Cell Biology
|April 18, 1998
Summary
The protein Eps15 interacts with the AP-2 complex, which is crucial for efficient endocytosis. Blocking this interaction inhibits the uptake of transferrin and epidermal growth factor, highlighting Eps15
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- The protein Eps15 is constitutively associated with the plasma membrane adaptor complex, AP-2.
- This association suggests a potential role for Eps15 in the process of endocytosis.
Purpose of the Study:
- To investigate the specific role of Eps15 in endocytosis.
- To elucidate the function of the AP-2/Eps15 association in cellular uptake processes.
Main Methods:
- Delineation of the Eps15 binding domain for AP-2.
- Expression of Eps15 COOH-terminal domain-green fluorescent protein (GFP) fusion proteins in HeLa cells.
- Cell-free assays using perforated A431 cells to study coated vesicle formation.
Main Results:
- Overexpression of Eps15 fusion protein with intact AP-2 binding sites inhibited transferrin endocytosis.
- A mutant Eps15 fusion protein lacking AP-2 binding sites had no effect on endocytosis.
- Eps15-derived fusion proteins inhibited both constitutive and ligand-induced endocytosis in cell-free assays.
Conclusions:
- The interaction between Eps15 and AP-2 is essential for efficient receptor-mediated endocytosis.
- Eps15 plays a critical role in the function of plasma membrane-coated pits.
- This study provides the first evidence for Eps15's involvement in endocytic pathways.