Lipid nanotechnologies for structural studies of membrane-associated proteins

Svetla Stoilova-McPhie, Kirill Grushin, Daniela Dalm, Jaimy Miller

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

We present a methodology of lipid nanotubes (LNT) and nanodisks technologies optimized in our laboratory for structural studies of membrane-associated proteins at close to physiological conditions. The application of these lipid nanotechnologies for structure determination by cryo-electron microscopy (cryo-EM) is fundamental for understanding and modulating their function. The LNTs in our studies are single bilayer galactosylceramide based nanotubes of ∼20 nm inner diameter and a few microns in length, that self-assemble in aqueous solutions. The lipid nanodisks (NDs) are self-assembled discoid lipid bilayers of ∼10 nm diameter, which are stabilized in aqueous solutions by a belt of amphipathic helical scaffold proteins. By combining LNT and ND technologies, we can examine structurally how the membrane curvature and lipid composition modulates the function of the membrane-associated proteins. As proof of principle, we have engineered these lipid nanotechnologies to mimic the activated platelet's phosphtaidylserine rich membrane and have successfully assembled functional membrane-bound coagulation factor VIII in vitro for structure determination by cryo-EM. The macromolecular organization of the proteins bound to ND and LNT are further defined by fitting the known atomic structures within the calculated three-dimensional maps. The combination of LNT and ND technologies offers a means to control the design and assembly of a wide range of functional membrane-associated proteins and complexes for structural studies by cryo-EM. The presented results confirm the suitability of the developed methodology for studying the functional structure of membrane-associated proteins, such as the coagulation factors, at a close to physiological environment.

Original languageEnglish (US)
Pages (from-to)2902-2909
Number of pages8
JournalProteins: Structure, Function and Bioinformatics
Volume82
Issue number11
DOIs
StatePublished - 2014

Keywords

  • Cryoelectron microscopy
  • Lipid nanodisks
  • Lipid nanotubes
  • Membrane-associated proteins
  • Protein structure

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology

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