Lipid nanotechnologies for structural studies of membrane-associated proteins

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

Research output: Contribution to journalArticle

6 Citations (Scopus)

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

Fingerprint

Nanotechnology
Nanotubes
Membrane Proteins
Lipids
Cryoelectron Microscopy
Electron microscopy
Technology
Galactosylceramides
Membranes
Lipid bilayers
Blood Coagulation Factors
Factor VIII
Lipid Bilayers
Membrane Lipids
Platelets
Scaffolds
Proteins
Blood Platelets
Chemical analysis

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Structural Biology
  • Molecular Biology

Cite this

Lipid nanotechnologies for structural studies of membrane-associated proteins. / Stoilova-McPhie, Svetla; Grushin, Kirill; Dalm, Daniela; Miller, Jaimy.

In: Proteins: Structure, Function and Bioinformatics, Vol. 82, No. 11, 2014, p. 2902-2909.

Research output: Contribution to journalArticle

Stoilova-McPhie, Svetla ; Grushin, Kirill ; Dalm, Daniela ; Miller, Jaimy. / Lipid nanotechnologies for structural studies of membrane-associated proteins. In: Proteins: Structure, Function and Bioinformatics. 2014 ; Vol. 82, No. 11. pp. 2902-2909.
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