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ProLIF - quantitative integrin protein-protein interactions and synergistic membrane effects on proteoliposomes. | LitMetric

AI Article Synopsis

  • Integrin receptors are essential for various biological processes by forming large protein complexes at their inner membrane interface, but existing methods to study these interactions have limitations, especially in realistic biological environments.
  • The new technique, termed ProLIF, allows researchers to analyze integrin interactions within liposomes using flow cytometry, providing a more accurate representation of how integrins function in a cell membrane context.
  • ProLIF simplifies the process by enabling the measurement of protein interactions directly from cell lysates without the need for prior purification, and it can accommodate high-throughput studies to explore complex membrane-dependent interactions.

Article Abstract

Integrin transmembrane receptors control a wide range of biological interactions by triggering the assembly of large multiprotein complexes at their cytoplasmic interface. Diverse methods have been used to investigate interactions between integrins and intracellular proteins, and predominantly include peptide-based pulldowns and biochemical immuno-isolations from detergent-solubilised cell lysates. However, quantitative methods to probe integrin-protein interactions in a more biologically relevant context where the integrin is embedded within a lipid bilayer have been lacking. Here, we describe 'protein-liposome interactions by flow cytometry' (denoted ProLIF), a technique to reconstitute recombinant integrin transmembrane domains (TMDs) and cytoplasmic tail (CT) fragments in liposomes as individual subunits or as αβ heterodimers and, via flow cytometry, allow rapid and quantitative measurement of protein interactions with these membrane-embedded integrins. Importantly, the assay can analyse binding of fluorescent proteins directly from cell lysates without further purification steps. Moreover, the effect of membrane composition, such as PI(4,5)P incorporation, on protein recruitment to the integrin CTs can be analysed. ProLIF requires no specific instrumentation and can be applied to measure a broad range of membrane-dependent protein-protein interactions with the potential for high-throughput/multiplex analyses.This article has associated First Person interviews with the first authors of the paper (see doi: 10.1242/jcs.223644 and doi: 10.1242/jcs.223719).

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6398480PMC
http://dx.doi.org/10.1242/jcs.214270DOI Listing

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