Use of dynasore, the small molecule inhibitor of dynamin, in the regulation of endocytosis.
The large GTPase dynamin is essential for clathrin-dependent coated-vesicle formation. Dynasore is a cell-permeable small molecule that inhibits the GTPase activity of dynamin1, dynamin2 and Drp1, the mitochondrial dynamin. Dynasore was discovered in a screen of approximately 16,000 compounds for inhibitors of the dynamin2 GTPase. Dynasore is a noncompetitive inhibitor of dynamin GTPase activity and blocks dynamin-dependent endocytosis in cells, including neurons. It is fast acting (seconds) and its inhibitory effect in cells can be reversed by washout. Here we present a detailed synthesis protocol for dynasore, and describe a series of experiments used to analyze the inhibitory effects of dynasore on dynamin in vitro and to study the effects of dynasore on endocytosis in cells.
Kirchhausen
Tom
T
Department of Cell Biology, Harvard Medical School, and IDI Immune Research Institute, Boston, Massachusetts, USA.
Macia
Eric
E
Pelish
Henry E
HE
D000818:Animals; D003599:Cytoskeleton / Q000187:drug effects; D034285:Dynamin II / Q000037:antagonists & inhibitors; D034281:Dynamins / Q000032:analysis / Q000037:antagonists & inhibitors*; D004705:Endocytosis; D006801:Humans; D006835:Hydrazones / Q000494:pharmacology*; D021381:Protein Transport / Q000187:drug effects; D018411:Spodoptera
D006835:Hydrazones; C511794:N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide; D034285:Dynamin II; D034281:Dynamins