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Updated: Feb 14, 2026

Co-Translational Insertion of Membrane Proteins into Preformed Nanodiscs
Published on: November 19, 2020
Production and Structural Analysis of Membrane-Anchored Proteins in Phospholipid Nanodiscs
Kolio Raltchev1,2, Joka Pipercevic1,2, Franz Hagn1,2
1Bavarian NMR Center at the Department of Chemistry and Institute for Advanced Study, Technical University of Munich, Lichtenbergstrasse 4, 85747, Garching, Germany.
Abstract:
Structural studies on membrane-anchored proteins containing a transmembrane (TM) helix have been hampered by difficulties in producing these proteins in a natively folded form. Detergents that are required to solubilize the hydrophobic TM helix usually destabilize the soluble domain. Thus, TM helices are removed for structural studies, which neglects the pivotal role of a membrane on protein function. This work presents a versatile strategy for the production of this protein class attached to phospholipid nanodiscs. By inserting the TM-helix into nanodiscs and a subsequent SortaseA-mediated ligation of the soluble domain, membrane-anchored BclxL could be obtained in a folded conformation. This strategy is suitable for high-resolution structure determination as well as for probing membrane location by NMR. This method will be applicable to a wide range of membrane-anchored proteins and will be useful to decipher their functional role in a native membrane environment.
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