Abstract
Membrane protein research suffers from the drawback that detergents, which are commonly used to solubilize integral membrane proteins (IMPs), often lead to protein instability and reduced activity. Recently, lipid nanodiscs (NDs) and saposin-lipoprotein particles (Salipro) have emerged as alternative carrier systems that keep membrane proteins in a native-like lipidic solution environment and are suitable for biophysical and structural studies. Here, we systematically compare nanodiscs and Salipros with respect to long-term stability as well as activity and stability of the incorporated membrane protein using the ABC transporter MsbA as model system. Our results show that both systems are suitable for activity measurements as well as structural studies in solution. Based on our results we suggest screening of different lipids with respect to activity and stability of the incorporated IMP before performing structural studies.
Acknowledgements
We thank Katharina Veith for technical assistance, Jens Frauenfeld (Salipro Biotech AB), Christian Löw and Franz Hagn for plasmids and advice/protocols and members of the Tidow lab for helpful discussions. This work is partially based upon experiments performed at the SANS-1 instrument operated by HZG and FRM II at the Heinz Maier-Leibnitz Zentrum (MLZ), Garching, Germany. The authors gratefully acknowledge the financial support provided by HZG to perform the neutron scattering measurements at the Heinz Maier-Leibnitz Zentrum (MLZ), Garching, Germany. We acknowledge access to the Sample Preparation and Characterization (SPC) Facility of EMBL, Hamburg. This work was funded by the Cluster of Excellence ‘The Hamburg Centre for Ultrafast Imaging’ of the Deutsche Forschungsgemeinschaft (DFG) – EXC 1074 – project ID 194651731.
Conflict of interest statement: The authors declare no competing financial interests.
References
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