CRAFTing Delivery of Membrane Proteins into Protocells using Nanodiscs

Stępień, Piotr; Świątek, Sylwia; Robles, Manuel Yamil Yusef; Markiewicz-Mizera, Joanna; Balakrishnan, Dhanasekaran; Inaba-Inoue, Satomi; De Vries, Alex H.; Beis, Konstantinos; Marrink, Siewert J.; Heddle, Jonathan G.

Abstract

For the successfulgenerativeengineeringof functionalartificialcells,a convenientandcontrollablemeansof deliveringmembraneproteinsinto membranelipidbilayersis necessary.Herewereporta deliverysystemthatachievesthis by employingmembraneprotein-carryingnanodiscsandthecalcium-dependentfusionofphosphatidylserinelipidmembranes.We showthat lipidnanodiscscanfuse a transportedlipidbilayerwith the lipidbilayersof smallunilamellarvesicles(SUVs)or giantunilamellarvesicles(GUVs)whileavoidingrecipientvesiclesaggregation.Thisis triggeredby a simple,transientincreasein calciumconcentration,whichresultsin efficientand rapidfusionin a one-potreaction.Furthermore,nanodiscscan be loadedwithmembraneproteinsthatcan be deliveredintotargetSUVor GUVmembranesin a detergent-independentfashionwhileretainingtheirfunctionality.Nanodiscshavea provenabilityto carrya widerangeof membraneproteins,controltheiroligomericstate,and arehighlyadaptable.Giventhis, our approachmay be the basisfor the developmentof usefultoolsthat will allowbespokedeliveryofmembraneproteinsto protocells,equippingthemwith the cell-likeabilityto exchangematerialacrossouter/subcellularmembranes.

Citation

Stępień, P., Świątek, S., Robles, M. Y. Y., Markiewicz-Mizera, J., Balakrishnan, D., Inaba-Inoue, S., …Heddle, J. G. (2023). CRAFTing Delivery of Membrane Proteins into Protocells using Nanodiscs. ACS Applied Materials and Interfaces, https://doi.org/10.1021/acsami.3c11894

• Journal Article Type: Article
• Acceptance Date: Nov 7, 2023
• Online Publication Date: Nov 28, 2023
• Publication Date: Nov 28, 2023
• Deposit Date: Dec 11, 2023
• Publicly Available Date: Dec 11, 2023
• Journal: ACS Applied Materials & Interfaces
• Print ISSN: 1944-8244
• Electronic ISSN: 1944-8252
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1021/acsami.3c11894
• Public URL: https://durham-repository.worktribe.com/output/2022442