Abstract
The solubilization and reconstitution of biological or liposomal membranes by detergents and biomolecules with detergent-like properties play a major role for technical applications (e.g., the isolation of membrane proteins) and biological phenomena (of, e.g., amphiphilic peptides). It is therefore important to know and understand the amounts of a given detergent required for the onset and completion of membrane solubilization and the detergent–lipid interactions in general. Lipid–detergent systems can form a variety of aggregate structures, which can be grouped into two pseudophases (lamellae and micelles) so that solubilization can be approximately described as a phase transition. Here we present a protocol for establishing the phase diagram and a detailed thermodynamic description of a lipid–detergent system based on isothermal titration calorimetry (ITC). The protocol can also be used to detect additive-induced membrane destabilization, permeabilization, domain formation and lipid-dependent transitions between rod-like and spherical micelles. A minimal protocol consisting of all sample preparation procedures and a single solubilization experiment can be accomplished within 2 days; a more extensive series comprising both solubilization and reconstitution experiments requires several days to a few weeks, depending on the number of titrations performed.
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Acknowledgements
This study was supported by grant 341920-07 from NSERC to H.H. and by grant KE 1478/1-1 from the Deutsche Forschungsgemeinschaft (DFG) to S.K.
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Heerklotz, H., Tsamaloukas, A. & Keller, S. Monitoring detergent-mediated solubilization and reconstitution of lipid membranes by isothermal titration calorimetry. Nat Protoc 4, 686–697 (2009). https://doi.org/10.1038/nprot.2009.35
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DOI: https://doi.org/10.1038/nprot.2009.35
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