ArticleNature protocols2016
A protocol for the systematic and quantitative measurement of protein-lipid interactions using the liposome-microarray-based assay.
Article in Nature protocols, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Exploring lipid-protein interactions in plant membranes.Journal of experimental botany · 2024Review
- The conformational plasticity of structurally unrelated lipid transport proteins correlates with their mode of action.PLoS biology · 2024Article
- Protein-Protein Interactions on Membrane Surfaces Analysed Using Pull-Downs with Supported Bilayers on Silica Beads.The Journal of membrane biology · 2022Article
- Encapsulated droplet interface bilayers as a platform for high-throughput membrane studies.Soft matter · 2022Article
- Protein Interaction Network Biology in Neuroscience.Proteomics · 2021Review
- Liposomes: Structure, Biomedical Applications, and Stability Parameters With Emphasis on Cholesterol.Frontiers in bioengineering and biotechnology · 2021Review
- Structures of three MORN repeat proteins and a re-evaluation of the proposed lipid-binding properties of MORN repeats.PloS one · 2020Article
- Perturbation of the yeast mitochondrial lipidome and associated membrane proteins following heterologous expression of Artemia-ANT.Scientific reports · 2018Article
- Mitochondria-specific photoactivation to monitor local sphingosine metabolism and function.eLife · 2018Article
- Critical size dependence of domain formation observed in coarse-grained simulations of bilayers composed of ternary lipid mixtures.The Journal of chemical physics · 2017Article
- Exploring functional membrane microdomains in bacteria: an overview.Current opinion in microbiology · 2017Review
- Phospholipid arrays on porous polymer coatings generated by micro-contact spotting.Beilstein journal of nanotechnology · 2017Article
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Authors and funding
9 authors.
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Abstract
Lipids organize the activity of the cell's proteome through a complex network of interactions. The assembly of comprehensive atlases embracing all protein-lipid interactions is an important challenge that requires innovative methods. We recently developed a liposome-microarray-based assay (LiMA) that integrates liposomes, microfluidics and fluorescence microscopy and which is capable of measuring protein recruitment to membranes in a quantitative and high-throughput manner. Compared with previous assays that are labor-intensive and difficult to scale up, LiMA improves the protein-lipid interaction assay throughput by at least three orders of magnitude. Here we provide a step-by-step LiMA protocol that includes the following: (i) the serial and generic production of the liposome microarray; (ii) its integration into a microfluidic format; (iii) the measurement of fluorescently labeled protein (either purified proteins or from cell lysate) recruitment to liposomal membranes using high-throughput microscopy; (iv) automated image analysis pipelines to quantify protein-lipid interactions; and (v) data quality analysis. In addition, we discuss the experimental design, including the relevant quality controls. Overall, the protocol-including device preparation, assay and data analysis-takes 6-8 d. This protocol paves the way for protein-lipid interaction screens to be performed on the proteome and lipidome scales.
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