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Antimycin A mimics a cell-death-inducing Bcl-2 homology domain 3

Nature Cell Biology volume 3, pages 183–191 (2001)Cite this article

Abstract

The Bcl-2-related survival proteins confer cellular resistance to a wide range of agents. Bcl-xL-expressing hepatocyte cell lines are resistant to tumour necrosis factor and anti-cancer drugs, but are more sensitive than isogenic control cells to antimycin A, an inhibitor of mitochondrial electron transfer. Computational molecular docking analysis predicted that antimycin A interacts with the Bcl-2 homology domain 3 (BH3)-binding hydrophobic groove of Bcl-xL. We demonstrate that antimycin A and a Bak BH3 peptide bind competitively to recombinant Bcl-2. Antimycin A and BH3 peptide both induce mitochondrial swelling and loss of ΔΨm on addition to mitochondria expressing Bcl-xL. The 2-methoxy derivative of antimycin A3 is inactive as an inhibitor of cellular respiration but still retains toxicity for Bcl-xL+ cells and mitochondria. Finally, antimycin A inhibits the pore-forming activity of Bcl-xL in synthetic liposomes, demonstrating that a small non-peptide ligand can directly inhibit the function of Bcl-2-related proteins.

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Figure 1: Bcl-xL expression confers paradoxical sensitivity to antimycin A.
Figure 2: Antimycin A induces Bcl-xL-dependent mitochondrial swelling and depolarization.
Figure 3: Effects of antimycin A on isolated mitochondria.
Figure 4: Molecular model of antimycin A bound to Bcl-xL.
Figure 5: Binding of antimycin A3 to Bcl-2 protein is inhibited by BH3 peptide.
Figure 6: Antimycin blocks Bcl-xL pore activity.
Figure 7: Binding and functional studies of antimycin A3 derivatives.

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Acknowledgements

We thank N. Fausto for providing TAMH, AML-12 and NMH cell lines; C. Thompson for providing FL5.12-Bcl-xL and FL5.12-neo cell lines; J. Foote and J. O'Neill for help and discussions; and R. Wiseman for assistance with oximetry measurements. This work was supported by NIH grant CA15704-26 (D.M.H.) and by an American Cancer Society grant RPG-97-173-01-LBC (K.Y.J.Z.). S.T. is an American Gastroenterological Association Research Scholar and a recipient of the Daland Fellowship for Research in Clinical Medicine from American Philosophical Society.

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Authors and Affiliations

  1. Division of Gastroenterology, Department of Medicine, University of Washington, Seattle, 98195, Washington, USA

    Shie-Pon Tzung

  2. Division of Basic Science, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N., Seattle, 98109, Washington, USA

    Kristine M. Kim & Kam Y. J. Zhang

  3. Division of Clinical Research, Fred Hutchinson Cancer Research Center, 1100 Fairview Ave N., Seattle, 98109, Washington, USA

    Chris D. Giedt, Julian Simon & David M. Hockenbery

  4. Laboratory of Cellular and Molecular Biophysics, National Institute of Child Health and Human Disease, National Institutes of Health , Bethesda, 20892, Maryland, USA

    Gorka Basañez & Joshua Zimmerberg

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Correspondence to David M. Hockenbery.

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Tzung, SP., Kim, K., Basañez, G. et al. Antimycin A mimics a cell-death-inducing Bcl-2 homology domain 3. Nat Cell Biol 3, 183–191 (2001). https://doi.org/10.1038/35055095

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