ArticleMethods in enzymology2019
Synthetic cell-like membrane interfaces for probing dynamic protein-lipid interactions.
Article in Methods in enzymology, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed.
- Article
- Initiation of lumen formation from junctions via differential actomyosin contractility regulated by dynamic recruitment of Rasip1.Nature communications · 2024Article
- Development and Application of an Optogenetic Manipulation System to Suppress Actomyosin Activity inInternational journal of molecular sciences · 2023Article
- Article
- Computational framework for single-cell spatiotemporal dynamics of optogenetic membrane recruitment.Cell reports methods · 2022Article
- Single-Component Optogenetic Tools for Inducible RhoA GTPase Signaling.Advanced biology · 2021Article
- Optogenetic Rac1 engineered from membrane lipid-binding RGS-LOV for inducible lamellipodia formation.Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology · 2020Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
The ability to rapidly screen interactions between proteins and membrane-like interfaces would aid in establishing the structure-function of protein-lipid interactions, provide a platform for engineering lipid-interacting protein tools, and potentially inform the signaling mechanisms and dynamics of membrane-associated proteins. Here, we describe the preparation and application of water-in-oil (w/o) emulsions with lipid-stabilized droplet interfaces that emulate the plasma membrane inner leaflet with tunable composition. Fluorescently labeled proteins are easily visualized in these synthetic cell-like droplets on an automated inverted fluorescence microscope, thus allowing for both rapid screening of relative binding and spatiotemporally resolved analyses of for example, protein-interface association and dissociation dynamics and competitive interactions, using commonplace instrumentation. We provide protocols for droplet formation, automated imaging assays and analysis, and the production of the positive control protein BcLOV4, a natural photoreceptor with a directly light-regulated interaction with anionic membrane phospholipids that is useful for optogenetic membrane recruitment.
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Registered trials
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