ArticleJournal of visualized experiments : JoVE2018
In Vitro Reconstitution of Self-Organizing Protein Patterns on Supported Lipid Bilayers.
Article in Journal of visualized experiments : JoVE, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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Who cites it
18 citing papers in PubMed.
- Article
- Cell-Size Confinement Drives Size-Dependent Scaling of Intracellular Reaction-Diffusion Waves for Robust Patterning.Small science · 2026Article
- 3D pattern formation of a protein-membrane suspension.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Self-organized spatial targeting of contractile actomyosin rings for synthetic cell division.Nature communications · 2024Article
- Freestanding bilayer microscope for single-molecule imaging of membrane proteins.Science advances · 2024Article
- Patterning of the cell cortex by Rho GTPases.Nature reviews. Molecular cell biology · 2024Review
- Machine learning-aided design and screening of an emergent protein function in synthetic cells.Nature communications · 2024Article
- MipZ caps the plus-end of FtsZ polymers to promote their rapid disassembly.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Reconstitution of Phase-Separated Signaling Clusters and Actin Polymerization on Supported Lipid Bilayers.Frontiers in cell and developmental biology · 2022Article
- Cardiolipin-Containing Lipid Membranes Attract the Bacterial Cell Division Protein DivIVA.International journal of molecular sciences · 2021Article
- Increasing MinD's Membrane Affinity Yields Standing Wave Oscillations and Functional Gradients on Flat Membranes.ACS synthetic biology · 2021Article
- Engineering spatiotemporal organization and dynamics in synthetic cells.Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology · 2021Review
- Probing Biomolecular Interactions by a Pattern-Forming Peptide-Conjugate Sensor.Bioconjugate chemistry · 2021Article
- Article
- The E. coli MinCDE system in the regulation of protein patterns and gradients.Cellular and molecular life sciences : CMLS · 2019Review
- Reconstitution and Coupling of DNA Replication and Segregation in a Biomimetic System.Chembiochem : a European journal of chemical biology · 2019Article
- Design of Sealable Custom-Shaped Cell Mimicries Based on Self-Assembled Monolayers on CYTOP Polymer.ACS applied materials & interfaces · 2019Article
- The MinDE system is a generic spatial cue for membrane protein distribution in vitro.Nature communications · 2018Article
Corrections and comments
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Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Many aspects of the fundamental spatiotemporal organization of cells are governed by reaction-diffusion type systems. In vitro reconstitution of such systems allows for detailed studies of their underlying mechanisms which would not be feasible in vivo. Here, we provide a protocol for the in vitro reconstitution of the MinCDE system of Escherichia coli, which positions the cell division septum in the cell middle. The assay is designed to supply only the components necessary for self-organization, namely a membrane, the two proteins MinD and MinE and energy in the form of ATP. We therefore fabricate an open reaction chamber on a coverslip, on which a supported lipid bilayer is formed. The open design of the chamber allows for optimal preparation of the lipid bilayer and controlled manipulation of the bulk content. The two proteins, MinD and MinE, as well as ATP, are then added into the bulk volume above the membrane. Imaging is possible by many optical microscopies, as the design supports confocal, wide-field and TIRF microscopy alike. In a variation of the protocol, the lipid bilayer is formed on a patterned support, on cell-shaped PDMS microstructures, instead of glass. Lowering the bulk solution to the rim of these compartments encloses the reaction in a smaller compartment and provides boundaries that allow mimicking of in vivo oscillatory behavior. Taken together, we describe protocols to reconstitute the MinCDE system both with and without spatial confinement, allowing researchers to precisely control all aspects influencing pattern formation, such as concentration ranges and addition of other factors or proteins, and to systematically increase system complexity in a relatively simple experimental setup.
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