Lipid nanotubes as substrates for helical crystallization of macromolecules

Elizabeth M. Wilson-Kubalek, Rhoderick E. Brown, Hervé Celia, Ronald A. Milligan

Research output: Contribution to journalArticlepeer-review

135 Scopus citations

Abstract

A general approach for crystallization of proteins in a fast and simple 'manner would be of immense interest to biologists studying protein structure-function relationships. Here, we describe a method that we have developed for promoting the formation of helical arrays of proteins and macromolecular assemblies. Electron micrographs of the arrays are suitable for helical image analysis and three-dimensional reconstruction. We show that hydrated mixtures of the glycolipid galactosylceramide (GalCer) and derivatized lipids or charged lipids form unilamellar nanotubules. The tubules bind proteins in a specific manner via high affinity ligands on the polar head groups of the lipid or via electrostatic interactions. By doping the GalCer with a novel nickel-containing lipid, we have been able to form helical arrays of two histidine-tagged proteins. Similarly, doping with a biotinylated lipid allows crystallization of streptavidin. Finally, three proteins with affinity for positively or negatively charged lipid layers formed helical arrays on appropriately charged tubules. The generality of this method may allow a wide variety of proteins to be crystallized on lipid nanotubes under physiological conditions.

Original languageEnglish (US)
Pages (from-to)8040-8045
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume95
Issue number14
DOIs
StatePublished - Jul 7 1998

Keywords

  • Electron microscopy
  • Galactosylceramides
  • Helical arrays
  • Tubules

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