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

Adam McCluskey, James A Daniel, Gordana Hadzic, Ngoc Chau, Emma L Clayton, Anna Mariana, Ainslie Whiting, Nick N Gorgani, Jonathan Lloyd, Annie Quan, Lia Moshkanbaryans, Sai Krishnan, Swetha Perera, Megan Chircop, Lisa von Kleist, Andrew B McGeachie, Mark T Howes, Robert G Parton, Michael Campbell, Jennette A SakoffXuefeng Wang, Jian-Yuan Sun, Mark J Robertson, Fiona M Deane, Tam H Nguyen, Frederic A Meunier, Michael A Cousin, Phillip J Robinson

Research output: Contribution to journalArticlepeer-review

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.
Original languageEnglish
Pages (from-to)1272-89
Number of pages18
JournalTraffic
Volume14
Issue number12
DOIs
Publication statusPublished - Dec 2013

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