Cycles of depalmitoylation and repalmitoylation critically control the steady-state localization and function of various peripheral membrane proteins, such as Ras proto-oncogene products. Interference with acylation using small molecules is a strategy to modulate cellular localization—and thereby unregulated signaling—caused by palmitoylated Ras proteins. We present the knowledge-based development and characterization of a potent inhibitor of acyl protein thioesterase 1 (APT1), a bona fide depalmitoylating enzyme that is, so far, poorly characterized in cells. The inhibitor, palmostatin B, perturbs the cellular acylation cycle at the level of depalmitoylation and thereby causes a loss of the precise steady-state localization of palmitoylated Ras. As a consequence, palmostatin B induces partial phenotypic reversion in oncogenic HRasG12V-transformed fibroblasts. We identify APT1 as one of the thioesterases in the acylation cycle and show that this protein is a cellular target of the inhibitor.
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We are grateful to H. Schütz for technical assistance with the western blots and immunofluorescence experiments. This work was financially supported by the Max-Planck Gesellschaft, the Deutsche Forschungsgemeinschaft, the Netherlands Organization for Scientific Research (NWO) (TALENT-stipendium for F.J.D.) and the European Union (Marie Curie fellowship for F.J.D.). N.V. is grateful to the International Max Planck Research School-Chemical Biology for providing a doctoral fellowship. C.H. and R.B. are grateful to the Alexander von Humboldt Foundation for postdoctoral fellowships. We thank the US National Institutes of Health for a Chemical/Biology Interface (CBI) Training Grant (to J.W.K.).
The authors declare no competing financial interests.
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Dekker, F., Rocks, O., Vartak, N. et al. Small-molecule inhibition of APT1 affects Ras localization and signaling. Nat Chem Biol 6, 449–456 (2010). https://doi.org/10.1038/nchembio.362
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