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Actin is the primary cellular receptor of bistramide A

Nature Chemical Biology volume 1, pages 383–388 (2005)Cite this article

Abstract

Bistramide A (1) is a marine natural product with broad, potent antiproliferative effects1,2,3,4,5,6,7. Bistramide A has been reported to selectively activate protein kinase C (PKC) δ, leading to the view that PKCδ is the principal mediator of antiproliferative activity of this natural product8. Contrary to this observation, we established that bistramide A binds PKCδ with low affinity, does not activate this kinase in vitro and does not translocate GFP-PKCδ. Furthermore, we identified actin as the cellular receptor of bistramide A. We report that bistramide A disrupts the actin cytoskeleton, inhibits actin polymerization, depolymerizes filamentous F-actin in vitro and binds directly to monomeric G-actin in a 1:1 ratio with a Kd of 7 nM. We also constructed a fully synthetic9 bistramide A–based affinity matrix and isolated actin as a specific bistramide A–binding protein. This activity provides a molecular explanation for the potent antiproliferative effects of bistramide A, identifying it as a new biochemical tool for studies of the actin cytoskeleton and as a potential lead for development of a new class of antitumor agents7,10.

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Figure 1
Figure 2: Effects of bistramide A on activation, binding and translocation of PKCδ.
Figure 3: Effect of bistramide A (bis A) on actin in A549 cells and in vitro.
Figure 4: Purification of bistramide A–binding proteins from A549 cell lysate.

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Acknowledgements

We thank G.F. Biard for a generous sample of natural bistramide A and Y. Lee for assistance with an affinity purification procedure. We also thank T. Meyer for imaging assistance and plasmids and D. Mochly-Rosen for binding assay support. Financial support was provided by the American Cancer Society (RSG-04-017-CDD) and the US National Institutes of Health (Grant CA31845). A.V.S. acknowledges the support of Burroughs Wellcome Fund Interfaces #1001774. J.L.B. is supported by an ACS Medicinal Chemistry Predoctoral Fellowship. S.A.K. thanks the Dreyfus Foundation for a Teacher-Scholar Award, Amgen for a New Investigator's Award and GlaxoSmithKline for a Chemistry Scholars Award. S.A.K. is a fellow of the Alfred P. Sloan Foundation.

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Authors and Affiliations

  1. Department of Chemistry, University of Chicago, Chicago, 60637, Illinois, USA

    Alexander V Statsuk & Sergey A Kozmin

  2. Division of Cancer Treatment and Diagnosis, Toxicology and Pharmacology Branch, Developmental Therapeutics Program, National Cancer Institute, National Institutes of Health, Frederick, 21702, Maryland, USA

    Ruoli Bai & Ernest Hamel

  3. Department of Chemistry and Department of Molecular Pharmacology, Stanford University, Stanford, 94305, California, USA

    Jeremy L Baryza, Vishal A Verma & Paul A Wender

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  1. Alexander V Statsuk
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Correspondence to Sergey A Kozmin.

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Supplementary information

Supplementary Fig. 1

Effect of bistramide A on cancer cell cycle. (PDF 382 kb)

Supplementary Fig. 2

Thermodynamic parameters of actin-latrunculin A binding. (PDF 349 kb)

Supplementary Fig. 3

Purification of bistramide A-binding proteins using affinity matrix. (PDF 181 kb)

Supplementary Table 1

Cancer cell growth and viability assays for bistramide A. (PDF 166 kb)

Supplementary Methods (PDF 94 kb)

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Statsuk, A., Bai, R., Baryza, J. et al. Actin is the primary cellular receptor of bistramide A. Nat Chem Biol 1, 383–388 (2005). https://doi.org/10.1038/nchembio748

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