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. 2022 May;289(10):2959-2970.
doi: 10.1111/febs.16327. Epub 2021 Dec 27.

Cryo-EM structure of MsbA in saposin-lipid nanoparticles (Salipro) provides insights into nucleotide coordination

Dominique-Maurice Kehlenbeck  1   2   3   4 , Daouda A K Traore  3   4   5   6 , Inokentijs Josts  1   2 , Simon Sander  1   2 , Martine Moulin  3   4 , Michael Haertlein  3   4 , Sylvain Prevost  7 , V Trevor Forsyth  3   4   5 , Henning Tidow  1
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Free article

Cryo-EM structure of MsbA in saposin-lipid nanoparticles (Salipro) provides insights into nucleotide coordination

Dominique-Maurice Kehlenbeck et al. FEBS J. 2022 May.
Free article

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Abstract

The ATP-binding cassette transporter MsbA is a lipid flippase, translocating lipid A, glycolipids, and lipopolysaccharides from the inner to the outer leaflet of the inner membrane of Gram-negative bacteria. It has been used as a model system for time-resolved structural studies as several MsbA structures in different states and reconstitution systems (detergent/nanodiscs/peptidiscs) are available. However, due to the limited resolution of the available structures, detailed structural information on the bound nucleotides has remained elusive. Here, we have reconstituted MsbA in saposin A-lipoprotein nanoparticles (Salipro) and determined the structure of ADP-vanadate-bound MsbA by single-particle cryo-electron microscopy to 3.5 Å resolution. This procedure has resulted in significantly improved resolution and enabled us to model all side chains and visualise detailed ADP-vanadate interactions in the nucleotide-binding domains. The approach may be applicable to other dynamic membrane proteins.

Keywords: MsbA; Salipro; cryo-EM structure; integral membrane proteins; saposin-lipoprotein nanoparticles.

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