Dynamin self-assembly stimulates its GTPase activity

J Biol Chem. 1996 Sep 13;271(37):22310-4. doi: 10.1074/jbc.271.37.22310.

Abstract

GTP hydrolysis by dynamin is required to drive coated vesicle budding at the plasma membrane. A diverse set of molecules including microtubules, grb2, and acidic phospholipids stimulate dynamin GTPase activity in vitro, although the physiological relevance of these effectors remains to be determined. Dynamin has been shown to assemble around microtubules, the most potent stimulatory molecule, into structures indistinguishable by electron microscopy from collars captured in vivo at the necks of endocytic coated pits. Under low ionic strength conditions purified dynamin self-assembles into rings and helical stacks of rings. Here we show that dynamin self-assembly stimulates its GTPase activity as much as 10-fold. Thus, we identify dynamin, itself, as the first effector of dynamin GTPase activity known to be physiologically relevant. Assembled dynamin's stimulated GTPase activity is not dependent on the direct interaction of high affinity GTP binding sites since a mutant defective in GTP binding and hydrolysis can coassemble with and stimulate GTP hydrolysis by wild-type dynamin. Finally, we find that GTP destabilizes assembled dynamin structures, suggesting that the activated rates of GTP hydrolysis reflect a continuing cycle of assembly, GTP hydrolysis, and disassembly.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Cell Line
  • Dynamins
  • Electrophoresis, Polyacrylamide Gel
  • GTP Phosphohydrolases / chemistry
  • GTP Phosphohydrolases / metabolism*
  • Guanosine 5'-O-(3-Thiotriphosphate) / pharmacology
  • Guanosine Diphosphate / pharmacology
  • Guanosine Triphosphate / metabolism
  • Humans
  • Hydrogen-Ion Concentration
  • Hydrolysis
  • Microtubules / metabolism*
  • Protein Conformation / drug effects
  • Substrate Specificity

Substances

  • Guanosine Diphosphate
  • Guanosine 5'-O-(3-Thiotriphosphate)
  • Guanosine Triphosphate
  • GTP Phosphohydrolases
  • Dynamins