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Isolation of F1-ATPase from the Parasitic Protist Trypanosoma brucei

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Published on: January 22, 2019

Pigment epithelium-derived factor binds to cell-surface F(1)-ATP synthase.

Luigi Notari¹, Naokatu Arakaki, David Mueller

¹Section of Protein Structure and Function, Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, NIH, Bethesda, MD, USA.

The FEBS Journal

|April 24, 2010

Summary

Pigment epithelium-derived factor (PEDF) binds to cell-surface F(1)-ATP synthase, inhibiting its activity. This interaction is key to PEDF's anti-angiogenic effects by blocking endothelial cell functions.

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Published on: April 5, 2018

Area of Science:

Biochemistry
Cell Biology
Molecular Medicine

Background:

Pigment epithelium-derived factor (PEDF) is a known inhibitor of angiogenesis and endothelial cell functions.
The specific cell-surface receptor for PEDF was previously unidentified.
F(1)F(o)-ATP synthase components have recently been implicated as cell-surface receptors.

Purpose of the Study:

To identify the protein that binds Pigment epithelium-derived factor (PEDF) on endothelial cell surfaces.
To investigate the direct binding interaction between PEDF and F(1)-ATP synthase.
To explore the functional consequences of PEDF binding to cell-surface F(1)-ATP synthase.

Main Methods:

Protein fingerprinting to identify PEDF-binding proteins.
Size-exclusion ultrafiltration and surface plasmon resonance to assess binding kinetics.
Antibody capture assays to confirm the presence of F(1)-ATP synthase in endothelial membranes.
Measurement of extracellular ATP synthesis in endothelial cells.

Main Results:

Protein fingerprinting identified a 60 kDa PEDF-binding protein as F(1)-ATP synthase beta-subunit.
Recombinant PEDF formed a complex with F(1)-ATP synthase, demonstrating specific and high-affinity binding.
PEDF binding to F(1)-ATP synthase inhibited its interaction with angiostatin and reduced extracellular ATP production.
Antibodies against F(1)-beta subunit captured PEDF-binding components in endothelial plasma membranes.

Conclusions:

Pigment epithelium-derived factor (PEDF) directly binds to cell-surface F(1)-ATP synthase.
PEDF acts as a ligand for endothelial cell-surface F(1)F(o)-ATP synthase.
PEDF-mediated inhibition of ATP synthase activity is a potential mechanism underlying its anti-angiogenic properties.