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Updated: Feb 22, 2026
Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions
Published on: February 10, 2022
Characterization of human Akt1-lipid interactions by label-free differential scanning fluorimetry
Obada Bahra1, Michal Grzybek2, Ünal Coskun3
1Center of Membrane Biochemistry and Lipid Research, University Hospital and Faculty of Medicine Carl Gustav Carus; TU Dresden, Dresden, Germany; Paul Langerhans Institute Dresden (PLID) of the Helmholtz Center Munich at the University Hospital Carl Gustav Carus and Faculty of Medicine of the TU Dresden, Dresden, Germany; German Center for Diabetes Research (DZD), Neuherberg, Germany.
Abstract:
Understanding the role of lipids in protein function is key to obtaining details about cellular signaling pathways. Among the various methods available, label-free differential scanning fluorimetry has several advantages. This technique uses the intrinsic fluorescence of tryptophan and tyrosine residues, requires minute sample amounts, enables rapid, high-throughput analysis, and allows direct comparison of protein-lipid interactions in lipid bilayers and with water-soluble lipid analogues. Here, we present a detailed protocol for monitoring the interaction between human full-length Akt1 and phosphatidylinositol-3,4,5-trisphosphate (PIP3), either in solution with short-chain PIP3 (4:0/4:0) or within membranes containing PIP3 (16:0/16:0). This versatile protocol can easily be adapted to study other protein-ligand interactions.
