Self-assembled monolayers improve protein distribution on holey carbon cryo-EM supports

Sci Rep. 2014 Nov 18:4:7084. doi: 10.1038/srep07084.

Abstract

Poor partitioning of macromolecules into the holes of holey carbon support grids frequently limits structural determination by single particle cryo-electron microscopy (cryo-EM). Here, we present a method to deposit, on gold-coated carbon grids, a self-assembled monolayer whose surface properties can be controlled by chemical modification. We demonstrate the utility of this approach to drive partitioning of ionotropic glutamate receptors into the holes, thereby enabling 3D structural analysis using cryo-EM methods.

Publication types

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

MeSH terms

  • Animals
  • Bacterial Proteins / chemistry*
  • Carbon / chemistry
  • Chaperonin 60 / chemistry*
  • Cryoelectron Microscopy / instrumentation*
  • GluK2 Kainate Receptor
  • Gold / chemistry
  • Porosity
  • Rats
  • Receptors, AMPA / chemistry*
  • Receptors, Kainic Acid / chemistry*
  • Wettability

Substances

  • Bacterial Proteins
  • Chaperonin 60
  • Receptors, AMPA
  • Receptors, Kainic Acid
  • Carbon
  • Gold
  • glutamate receptor ionotropic, AMPA 2