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Serendipitous alkylation of a Plk1 ligand uncovers a new binding channel

Nature Chemical Biology volume 7, pages 595–601 (2011)Cite this article

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Abstract

We obtained unanticipated synthetic byproducts from alkylation of the δ1 nitrogen (N3) of the histidine imidazole ring of the polo-like kinase-1 (Plk1) polo-box domain (PBD)-binding peptide PLHSpT. For the highest-affinity byproduct, bearing a C6H5(CH2)8– group, a Plk1 PBD cocrystal structure revealed a new binding channel that had previously been occluded. An N-terminal PEGylated version of this peptide containing a hydrolytically stable phosphothreonyl residue (pT) bound the Plk1 PBD with affinity equal to that of the non-PEGylated parent but showed markedly less interaction with the PBDs of the two closely related proteins Plk2 and Plk3. Treatment of cultured cells with this PEGylated peptide resulted in delocalization of Plk1 from centrosomes and kinetochores and in chromosome misalignment that effectively induced mitotic block and apoptotic cell death. This work provides insights that might advance efforts to develop Plk1 PBD-binding inhibitors as potential Plk1-specific anticancer agents.

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Figure 1: X-ray co-crystal structures of Plk1 PBD complexed with peptides 1 and 4j.
Figure 2: Specific inhibition of the function of Plk1 PBD by peptide 6.

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Acknowledgements

This work was supported in part by grants ZIA BC 006198 (T.R.B.) and ZIA BC 010681 (K.S.L.) of the Intramural Research Program of the US National Institutes of Health, Center for Cancer Research, National Cancer Institute–Frederick and the US National Cancer Institute, US National Institutes of Health (F.L., J.-E.P., W.-J.Q., K.S.L. and T.R.B.), US National Institutes of Health grants GM60594, GM68762 and CA112967 (M.B.Y.) and the Deutsche Forschungsgemeinschaft (grant BE 4572/1-1) (T.B.). We thank M. Dyba of the Biophysics Resource of the Structural Biophysics Laboratory, National Cancer Institute Frederick for assistance in performing tandem MS studies and W. Dai, New York University School of Medicine for the Flag-Plk3 construct. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial product or organizations imply endorsement by the US government.

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  1. Fa Liu and Jung-Eun Park: These authors contributed equally to this work.

Authors and Affiliations

  1. Chemical Biology Laboratory, Molecular Discovery Program, Center for Cancer Research, National Cancer Institute-Frederick, Frederick, Maryland, USA

    Fa Liu, Wen-Jian Qian & Terrence R Burke Jr

  2. Laboratory of Metabolism, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA

    Jung-Eun Park & Kyung S Lee

  3. Department of Biology and Biological Engineering, Center for Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA

    Dan Lim & Michael B Yaffe

  4. Institute of Organic Chemistry, University of Leipzig, Leipzig, Germany

    Martin Gräber & Thorsten Berg

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Contributions

F.L., J.-E.P., W.-J.Q., D.L., M.G., T.B., M.B.Y., K.S.L. and T.R.B. designed the experiments; F.L., J.-E.P., W.-J.Q., D.L. and M.G. conducted the experiments and F.L., J.-E.P., W.-J.Q., D.L., M.G., T.B., M.B.Y., K.S.L. and T.R.B. analyzed the data and wrote the paper.

Corresponding authors

Correspondence to Kyung S Lee or Terrence R Burke Jr.

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Liu, F., Park, JE., Qian, WJ. et al. Serendipitous alkylation of a Plk1 ligand uncovers a new binding channel. Nat Chem Biol 7, 595–601 (2011). https://doi.org/10.1038/nchembio.614

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