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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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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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)
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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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DOI: https://doi.org/10.1038/nchembio748
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