Protocol for building synthetic protocell membranes that sense redox using synthetic phospholipids and natural lipids

• Published in: STAR protocols 2024-09, Vol.5 (3), p.103169, Article 103169
• Main Authors: Wang, Liming, Dresel, Mark J., Robinson, Monique, Izgu, Enver Cagri
• Format: Article
• Language: English
• Subjects: Artificial Cells - chemistry; Artificial Cells - metabolism; Chemistry; Hydrogen Peroxide - chemistry; Lipids - chemistry; Molecular/Chemical Probes; Oxidation-Reduction; Phospholipids - chemistry; Protocol; Unilamellar Liposomes - chemistry; Unilamellar Liposomes - metabolism
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.103169

Sensing is a critical function of artificial cells; however, this is challenging to realize using bottom-up approaches. Here, we present a protocol for building protocell membranes that sense cues important for redox biochemistry and signaling by combining synthetic phospholipids and natural lipids. We detail procedures for building giant unilamellar vesicles as protocell models that fluoresce in response to the biologically significant redox agents peroxynitrite, hydrogen peroxide, and hydrogen sulfide. For complete details on the use and execution of this protocol, please refer to (i) Gutierrez and Aggarwal et al.1 as well as (ii) Erguven and Wang et al.2 [Display omitted] •Preparation of diverse synthetic lipids with modular designs from a common precursor•Generation of giant unilamellar vesicles (GUVs) with chemically functional lipids•Demonstration by confocal imaging of how GUVs appear before and after redox treatment Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Sensing is a critical function of artificial cells; however, this is challenging to realize using bottom-up approaches. Here, we present a protocol for building protocell membranes that sense cues important for redox biochemistry and signaling by combining synthetic phospholipids and natural lipids. We detail procedures for building giant unilamellar vesicles as protocell models that fluoresce in response to the biologically significant redox agents peroxynitrite, hydrogen peroxide, and hydrogen sulfide.