Use of dynasore, the small molecule inhibitor of dynamin, in the regulation of endocytosis

Methods Enzymol. 2008;438:77-93. doi: 10.1016/S0076-6879(07)38006-3.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cytoskeleton / drug effects
  • Dynamin II / antagonists & inhibitors
  • Dynamins / analysis
  • Dynamins / antagonists & inhibitors*
  • Endocytosis
  • Humans
  • Hydrazones / pharmacology*
  • Protein Transport / drug effects
  • Spodoptera

Substances

  • Hydrazones
  • N'-(3,4-dihydroxybenzylidene)-3-hydroxy-2-naphthahydrazide
  • Dynamin II
  • Dynamins