These authors contributed equally to this work.
Building a Better Dynasore: The Dyngo Compounds Potently Inhibit Dynamin and Endocytosis†
Article first published online: 9 OCT 2013
© 2013 The Authors. Traffic Published by John Wiley & Sons Ltd
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Volume 14, Issue 12, pages 1272–1289, December 2013
How to Cite
McCluskey, A., Daniel, J. A., Hadzic, G., Chau, N., Clayton, E. L., Mariana, A., Whiting, A., Gorgani, N. N., Lloyd, J., Quan, A., Moshkanbaryans, L., Krishnan, S., Perera, S., Chircop, M., von Kleist, L., McGeachie, A. B., Howes, M. T., Parton, R. G., Campbell, M., Sakoff, J. A., Wang, X., Sun, J.-Y., Robertson, M. J., Deane, F. M., Nguyen, T. H., Meunier, F. A., Cousin, M. A. and Robinson, P. J. (2013), Building a Better Dynasore: The Dyngo Compounds Potently Inhibit Dynamin and Endocytosis. Traffic, 14: 1272–1289. doi: 10.1111/tra.12119
The copyright of this article has now been changed to open access since first published on 09 October 2013.
- Issue published online: 11 NOV 2013
- Article first published online: 9 OCT 2013
- Accepted manuscript online: 11 SEP 2013 12:49PM EST
- Manuscript Accepted: 11 SEP 2013
- Manuscript Revised: 9 SEP 2013
- Manuscript Received: 23 OCT 2012
- National Health and Medical Research Council (Australia)
- Epilepsy Research USA
- The Wellcome Trust. Grant Number: 084277
- The Children's Medical Research Institute
- Newcastle Innovation
- Ramaciotti Foundation
- Australian Cancer Research Foundation
- Ian Potter Foundation
- bulk endocytosis;
- drug discovery;
- high-throughput screening;
- small-molecule inhibitors;
- synaptic vesicle endocytosis
Dynamin GTPase activity increases when it oligomerizes either into helices in the presence of lipid templates or into rings in the presence of SH3 domain proteins. Dynasore is a dynamin inhibitor of moderate potency (IC50 ˜ 15 μM in vitro). We show that dynasore binds stoichiometrically to detergents used for in vitro drug screening, drastically reducing its potency (IC50 = 479 μM) and research tool utility. We synthesized a focused set of dihydroxyl and trihydroxyl dynasore analogs called the Dyngo™ compounds, five of which had improved potency, reduced detergent binding and reduced cytotoxicity, conferred by changes in the position and/or number of hydroxyl substituents. The Dyngo compound 4a was the most potent compound, exhibiting a 37-fold improvement in potency over dynasore for liposome-stimulated helical dynamin activity. In contrast, while dynasore about equally inhibited dynamin assembled in its helical or ring states, 4a and 6a exhibited >36-fold reduced activity against rings, suggesting that they can discriminate between helical or ring oligomerization states. 4a and 6a inhibited dynamin-dependent endocytosis of transferrin in multiple cell types (IC50 of 5.7 and 5.8 μM, respectively), at least sixfold more potently than dynasore, but had no effect on dynamin-independent endocytosis of cholera toxin. 4a also reduced synaptic vesicle endocytosis and activity-dependent bulk endocytosis in cultured neurons and synaptosomes. Overall, 4a and 6a are improved and versatile helical dynamin and endocytosis inhibitors in terms of potency, non-specific binding and cytotoxicity. The data further suggest that the ring oligomerization state of dynamin is not required for clathrin-mediated endocytosis.