Structural and functional studies of membrane remodeling machines

Methods Cell Biol. 2015;128:165-200. doi: 10.1016/bs.mcb.2015.02.007. Epub 2015 Apr 8.

Abstract

Building cells from their component parts will hinge upon our ability to reconstitute biochemical compartmentalization and exchange between membrane-delimited organelles. By contrast with our understanding of other cellular events, the mechanisms that govern membrane trafficking has lagged because the presence of phospholipid bilayers complicates the use of standard methods. This chapter describes in vitro methods for purifying, reconstituting, and visualizing membrane remodeling activities directly by electron cryomicroscopy.

Keywords: Bilayer; Dynamin; Electron cryomicroscopy; Endocytosis; Endophilin; Lipid-binding; Membrane remodeling; Mitochondria; cryo-EM.

Publication types

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

MeSH terms

  • Acyltransferases / metabolism*
  • Animals
  • Cell Line
  • Cell Membrane / metabolism*
  • Cryoelectron Microscopy / methods*
  • Dynamins / biosynthesis
  • Dynamins / metabolism*
  • Endocytosis / physiology
  • Escherichia coli / metabolism
  • Lipid Bilayers
  • Protein Structure, Tertiary
  • Sf9 Cells
  • Spodoptera

Substances

  • Lipid Bilayers
  • Acyltransferases
  • 2-acylglycerophosphate acyltransferase
  • Dynamins