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| Home > Publications database > Membrane protein scaffold systems and SAXS characterization |
| Abstract | PUBDB-2018-05607 |
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2018
Abstract: Infrastructure needed for the solution characterization of reconstituted membrane proteins (MPs) has been designed and implemented at the iNEXT partner synchrotron facilities Petra-III (P12, EMBL-HH) and SOLEIL (SWING, SOLEIL). HT-SEC-SAXS systems are now optimized for purification and screening and rapid structural data collection of MPs. Feasibility studies, testing translational targets and the analysis of several MPs reconstituted in detergents have been conducted, including the newly developed saposinlipid particles. An exhaustive assessment of the ability of the human saposin (Sap) proteins (A to D) to form nanoparticles (SapNPs) has been conducted including screening against a library of different detergent-solubilized lipids and lipid mixtures [1-2]. The SEC-SAXS data obtained demonstrates the typical lipid bilayer particle structure and modularity of the SapNPs. The mechanosensitive channel protein T2 has been successfully reconstituted in SapNPs and characterized by SAXS [2]. Work conducted using this infrastructure demonstrates the ready characterization of detergent solubilsed MPs and facilitated the establishment of generic protocols to be used as starting point to reconstitute any membrane protein in SapNPs. In addition, the work conducted in has provided the basis for further development on the automation of this HT-SEC-SAXS infrastructure. References [1] - J.A. Lyons, A. Bøggild, P. Nissen, J. Frauenfeld, Saposin-Lipoprotein Scaffolds for Structure Determination of Membrane Transporters. Methods in Enzymology, volume 594: A Structure-Function Toolbox for Membrane Transporter and Channels. 2017. p. 85-99. [2] - A. Flayhan, H.D.T. Mertens, Y.U. Blimke, M.M. Molledo, D. Svergun, C. Löw, Saposin lipid nanoparticles: a highly versatile and modular tool for membrane protein research. Structure, 26 1-11 2018.
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