Two-dimensional crystallization of integral membrane proteins for electron crystallography

Methods Mol Biol. 2010:654:187-205. doi: 10.1007/978-1-60761-762-4_10.

Abstract

Although membrane proteins make up 30% of the proteome and are a common target for therapeutic drugs, determination of their atomic structure remains a technical challenge. Electron crystallography represents an alternative to the conventional methods of X-ray diffraction and NMR and relies on the formation of two-dimensional crystals. These crystals are produced by reconstituting purified, detergent-solubilized membrane proteins back into the native environment of a lipid bilayer. This chapter reviews methods for producing two-dimensional crystals and for screening them by negative stain electron microscopy. In addition, we show examples of the different morphologies that are commonly obtained and describe basic image analysis procedures that can be used to evaluate their promise for structure determination by cryoelectron microscopy.

Publication types

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

MeSH terms

  • Cryoelectron Microscopy
  • Crystallography / methods*
  • Membrane Proteins / chemistry*
  • Membrane Proteins / ultrastructure
  • Microscopy, Electron

Substances

  • Membrane Proteins