Protein-lipid interactions

Here the authors detail the protocol for their recently developed liposome-microarray-based assay (LiMA) that integrates liposomes, microfluidics and fluorescence microscopy to enable the systematic and quantitative measurement of protein–lipid interactions.

Aqueous phase protein-lipid interactions have a role in cell physiology. This protocol describes the analysis of in vivo–assembled protein-lipid complexes in S. cerevisiae using TAP labels, size-exclusion chromatography, SDS-PAGE and HPTLC.

Genetic interactions reveal molecular and functional gene networks. Here systematic siRNA-mediated pairwise gene knockdown combined with high-content imaging and computational analysis measures genetic interactions at high throughput in human cells.

Lipid rafts are relatively ordered membrane domains that are enriched in cholesterol and saturated lipids, and selectively recruit other lipids and proteins. They are dynamic and heterogeneous in composition and are thus challenging to visualizein vivo. New technologies are providing novel insights into the formation, organization and functions of these membrane domains.

Lipids tailor membrane identities and function as molecular hubs in all cellular processes. The development of pioneering technologies, including affinity-purification lipidomics and the liposome microarray-based assay (LiMA), will enable researchers to decipher protein–lipid interactions and enhance our understanding of how lipids modulate protein function and structure.