Machin, J. M., Ranson, N. A. and Radford, S. E. orcid.org/0000-0002-3079-8039 (Accepted: 2025) Protein-induced membrane asymmetry modulates OMP folding kinetics and stability. Faraday Discussions. ISSN 1359-6640 (In Press)
Abstract
Biological membranes are asymmetric structures, with asymmetry arising from differences in lipid identity in each leaflet of the bilayer, as well as non-uniform distribution of lipids and small molecules in the membrane. Proteins can also induce and modulate membrane asymmetry based on their shape, sequence and interactions with lipids. How membrane asymmetry affects macromolecular behaviour is poorly understood because of the complexity of natural membrane systems, and difficulties in creating relevant asymmetric bilayer systems in vitro. Here, we present a method exploiting the efficient, unidirectional folding of the transmembrane β-barrel outer membrane protein, OmpA, to create asymmetric proteoliposomes with proteininduced dipoles of known direction (arising from sequence variation engineered into the OmpA loops). We then characterise the folding kinetics and stability of different OmpA variants into these proteoliposomes. We find that both the primary sequence of the folding OmpA and the dipole of the membrane into which folding occurs, play an important role for modulating the rate of folding. Critically, we find that by complementarily matching the charge on the folding protein to the membrane dipole it is possible to enhance both the folding kinetics and the stability of the folded OmpA. The results hint at how cells might exploit loop charge in membrane-embedded proteins to manipulate membrane environments for adaptation and survival.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | This is an author produced version of an article accepted for publication in Faraday Discussions, made available under the terms of the Creative Commons Attribution License (CC-BY), which permits unrestricted use, distribution and reproduction in any medium, provided the original work is properly cited. |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Molecular and Cellular Biology (Leeds) > Structural Molecular Biology (Leeds) |
Funding Information: | Funder Grant number Wellcome Trust 208385/Z/17/Z MRC (Medical Research Council) MR/Y012453/1 Royal Society RSRP\R1\211057 |
Depositing User: | Symplectic Publications |
Date Deposited: | 10 Jan 2025 09:48 |
Last Modified: | 27 Mar 2025 09:08 |
Status: | In Press |
Publisher: | RSC |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:221591 |