A Versatile Suspended Lipid Membrane System for Probing Membrane Remodeling and Disruption

• DOI: 10.20944/preprints202209.0119.v1
• Publication Date: 2022-09-09T06:36:47Z
• Authors: Achinta Sannigrahi; Vishwesh Harichar...; Muhsin Vannan Chalil; Debayani Chakraborty; Subrat Kumar Meher; Rahul Roy
• Keywords: 0301 basic medicine; 0303 health sciences; pore formation; Chemical technology; TP1-1185; Article; 3. Good health; virus fusion; 03 medical and health sciences; Chemical engineering; suspended bilayer; pore-forming toxin; Cytolysin A; virus fusion; pore formation; biophysics; TP155-156; suspended bilayer; pore-forming toxin; Cytolysin A

Artificial membrane systems can serve as models to investigate molecular mechanisms of different cellular processes, including transport, pore formation, and viral fusion. However, the current simulacrums such as SUVs, GUVs, and the supported lipid bilayers suffer from issues, namely high curvature, heterogeneity, and surface artefacts, respectively. Freestanding membranes provide a facile solution to these issues, but current systems developed by various groups use silicon or aluminium oxide wafers for fabrication that involves access to a dedicated nanolithography facility and high cost while conferring poor membrane stability. Here, we report the development, characterization and applications of an easy-to-fabricate suspended lipid bilayer (SULB) membrane platform leveraging commercial track-etched porous filters (PCTE) with defined microwell size. Our SULB system offers a platform to study the lipid composition-dependent structural and functional properties of membranes with exceptional stability in a high throughput fashion. With dye entrapped in PCTE microwells by SULB, we show that sphingomyelin significantly augments the activity of pore-forming toxin, Cytolysin A (ClyA) and the pore formation induces lipid exchange between the bilayer leaflets. Further, we demonstrate high efficiency and rapid kinetics of membrane fusion by dengue virus in our SULB platform. Our suspended bilayer membrane mimetic offers a novel platform to investigate a large class of biomembrane interactions and processes.