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Vol. 95, Issue 14, 8040-8045, July 7, 1998

Biophysics
 Lipid nanotubes as substrates for helical crystallization of macromolecules

(tubules/helical arrays/galactosylceramides/electron microscopy)

Elizabeth M. Wilson-Kubalek*, Rhoderick E. Brown*,dagger , Hervé Celia*, and Ronald A. Milligan*,Dagger

* Department of Cell Biology, MB25, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037; and dagger  The Hormel Institute, University of Minnesota, Austin, MN 55912

Edited by Roger D. Kornberg, Stanford University School of Medicine, Stanford, CA, and approved May 1, 1998 (received for review April 2, 1998)

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.

Dagger    To whom reprint requests should be addressed. e-mail: milligan{at}scripps.edu.

Copyright © 1998 by The National Academy of Sciences  0027-8424/98/958040-6$2.00/0
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