2016
DOI: 10.1038/nprot.2016.059
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A protocol for the systematic and quantitative measurement of protein–lipid interactions using the liposome-microarray-based assay

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 a… Show more

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Cited by 29 publications
(23 citation statements)
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“…LiMA enables sampling of a wide range of liposome compositions and curvatures in a single experiment. Readout is via the detection of two fluorophores - one on a carrier lipid in the liposomes, and one on the protein of interest (Saliba, Vonkova et al, 2014, Saliba, Vonkova et al, 2016). Colocalised signals are regarded as positive hits.…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…LiMA enables sampling of a wide range of liposome compositions and curvatures in a single experiment. Readout is via the detection of two fluorophores - one on a carrier lipid in the liposomes, and one on the protein of interest (Saliba, Vonkova et al, 2014, Saliba, Vonkova et al, 2016). Colocalised signals are regarded as positive hits.…”
Section: Resultsmentioning
confidence: 99%
“…LiMA (Saliba et al, 2014) was performed in the lab of Anne-Claude Gavin (EMBL Heidelberg, Germany) according to the standard protocol (Saliba et al, 2016). TbMORN1(2-15) tagged N-terminally with EGFP and two positive controls, PLCδ1-PH and Lactadherin-C2, both fused to superfolder GFP (sfGFP), were applied to microarrays printed with different signalling lipids.…”
Section: Methodsmentioning
confidence: 99%
“…In the last years some new techniques were introduced to explore lipid metabolism ( Wenk, 2005 ; da Silveira Dos Santos et al, 2014 ; Papan et al, 2014 ) and to map lipid-protein interactions ( Haberkant et al, 2016 ; Saliba et al, 2016 ). While proven to be valuable tools, they do not permit the acute manipulation of lipid amounts in living cells.…”
Section: Introductionmentioning
confidence: 99%
“…This method of preparation of liposomes is based on the microfluidization procedure employing a microfluidizer ( Figure 5) without using potentially toxic solvents (Mirakabad et al 2016, YuJi and. Microfluidizer has been applied in the pharmaceutical industry to produce liposomal products and pharmaceutical emulsions (Saliba et al 2016). This technique uses a divided pressure stream into passing each part across a tiny aperture and leading the flows to each other inside the chamber of microfluidizer (DevrimKara et al 2016).…”
Section: Microfluidizationmentioning
confidence: 99%