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Bahra, O. ; Grzybek, M.* ; Coskun, Ü.

Characterization of human Akt1-lipid interactions by label-free differential scanning fluorimetry.

In:. Elsevier, 2025. DOI: 10.1016/bs.mie.2025.11.002 (Methods Enzymol.)
DOI
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.
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Publication type Article: Edited volume or book chapter
Keywords Differential Scanning Fluorimetry ; Protein Lipid Interaction ; Protein Membrane Interaction ; Thermal Unfolding ; Tryptophan Fluorescence
ISSN (print) / ISBN 0076-6879
e-ISSN 0076-6879
Journal Methods in Enzymology
Publisher Elsevier
Reviewing status Peer reviewed
Institute(s) Institute of Pancreatic Islet Research (IPI)