Oligomerization state of dynamin 2 in cell membranes using TIRF and number and brightness analysis

Biophys J. 2011 Feb 2;100(3):L15-L17. doi: 10.1016/j.bpj.2010.12.3703.

Abstract

Dynamin 2 is an ubiquitously expressed ∼100 kDa GTPase involved in receptor-mediated endocytosis, Golgi budding, and cytoskeletal reorganization. Dynamin molecules assemble around the necks of budding vesicles and constrict membranes in a GTP-dependent process, resulting in vesicle release. The oligomerization state of dynamin 2 in the membrane is still controversial. We investigated dynamin 2 within the plasma membrane of live cells using total internal reflection microscopy coupled with number and brightness analysis. Our results demonstrate that dynamin 2 is primarily tetrameric throughout the entire cell membrane, aside from punctate structures that may correspond to regions of membrane vesiculation.

Publication types

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

MeSH terms

  • Animals
  • Cell Membrane / metabolism*
  • Dynamin II / chemistry*
  • Dynamin II / metabolism*
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism*
  • Green Fluorescent Proteins / metabolism
  • Mice
  • Microscopy, Fluorescence / methods*
  • Protein Structure, Quaternary
  • Rats
  • Recombinant Fusion Proteins / metabolism

Substances

  • Recombinant Fusion Proteins
  • enhanced green fluorescent protein
  • Green Fluorescent Proteins
  • Dynamin II