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Nat Methods. 2016 Apr;13(4):345-51. doi: 10.1038/nmeth.3801. Epub 2016 Mar 7.

A saposin-lipoprotein nanoparticle system for membrane proteins.

Author information

1
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
2
Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
3
Keck Advanced Microscopy Laboratory, Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, California, USA.
4
Structural and Computational Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, Germany.
5
Molecular Medicine Partnership Unit, European Molecular Biology Laboratory-Universitätsklinikum Heidelberg, Heidelberg, Germany.
6
School of Technology and Health, Royal Institute of Technology, Novum, Huddinge, Sweden.
7
EMBL Hamburg, Hamburg, Germany.
8
Howard Hughes Medical Institute, University of California San Francisco, San Francisco, California, USA.

Abstract

A limiting factor in membrane protein research is the ability to solubilize and stabilize such proteins. Detergents are used most often for solubilizing membrane proteins, but they are associated with protein instability and poor compatibility with structural and biophysical studies. Here we present a saposin-lipoprotein nanoparticle system, Salipro, which allows for the reconstitution of membrane proteins in a lipid environment that is stabilized by a scaffold of saposin proteins. We demonstrate the applicability of the method on two purified membrane protein complexes as well as by the direct solubilization and nanoparticle incorporation of a viral membrane protein complex from the virus membrane. Our approach facilitated high-resolution structural studies of the bacterial peptide transporter PeptTSo2 by single-particle cryo-electron microscopy (cryo-EM) and allowed us to stabilize the HIV envelope glycoprotein in a functional state.

PMID:
26950744
PMCID:
PMC4894539
DOI:
10.1038/nmeth.3801
[Indexed for MEDLINE]
Free PMC Article

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