Building a better dynasore: the dyngo compounds potently inhibit dynamin and endocytosis

McCluskey, Adam, Daniel, James A., Hadzic, Gordana, Chau, Ngoc , Clayton, Emma L., Mariana, Anna, Whiting, Ainslie, Gorgani, Nick N., Lloyd, Jonathan, Quan, Annie, Moshkanbaryans, Lia, Krishnan, Sai, Perera, Swetha, Chircop, Megan, von Kleist, Lisa, McGeachie, Andrew B., Howes, Mark T., Parton, Robert G., Campbell, Michael, Sakoff, Jennette A., Wang, Xuefeng, Sun, Jian-Yuan, Robertson, Mark J., Deane, Fiona M., Nguyen, Tam H., Meunier, Frederic A., Cousin, Michael A., and Robinson, Phillip J. (2013) Building a better dynasore: the dyngo compounds potently inhibit dynamin and endocytosis. Traffic, 14 (12). pp. 1272-1289.

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Abstract

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.

Item ID: 36832
Item Type: Article (Research - C1)
ISSN: 1600-0854
Keywords: bulk endocytosis, drug discovery, dynamin, high-throughput screening, small-molecule inhibitors, synaptic vesicle endocytosis
Additional Information:

Copyright © 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.

Funders: National Health and Medical Research Council of Australia (NHMRC), Epilepsy Research USA, Wellcome Trust, Children's Medical Research Institute, Newcastle Innovation, Ramaciotti Foundation, Australian Cancer Research Foundation, Ian Potter Foundation
Projects and Grants: Wellcome Trust (084277)
Date Deposited: 10 Dec 2014 07:31
FoR Codes: 03 CHEMICAL SCIENCES > 0305 Organic Chemistry > 030503 Organic Chemical Synthesis @ 100%
SEO Codes: 97 EXPANDING KNOWLEDGE > 970103 Expanding Knowledge in the Chemical Sciences @ 100%
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