Jens Frauenfeld, Robin Löving, Jean-Paul Armache, Andreas F-P Sonnen, Fatma Guettou, Per Moberg, Lin Zhu, Caroline Jegerschöld, Ali Flayhan, John A G Briggs, Henrik Garoff, Christian Löw, Yifan Cheng & Pär Nordlund
2016 vol: 13 pp: 345–351 doi:10.1038/nmeth.3801
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.
Topics: Detergents, Lipids, Membrane proteins, Supplements, Prometheus – nanoDSF, Proteins, Publications