Abstract
The ability to bind to and translocate across lipid bilayers is of paramount importance for the extracellular administration of intracellularly active compounds in cell biology, medicinal chemistry, and drug development. A combination of the so-called uptake and release experiments performed by high-sensitivity isothermal titration calorimetry provides a powerful and universally applicable tool for measuring membrane binding and translocation of various compound classes in a label-free manner in solution. The protocol presented here is designed for a quantitative analysis of microcalorimetric uptake and release titrations. In contrast with simpler approaches described previously, it is applicable also to electrically charged solutes, such as peptides and proteins, experimentally and clinically relevant surfactants, drugs, metal ions, and other ionic compounds.
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References
Seelig J (2004) Thermodynamics of lipid–peptide interactions. Biochim Biophys Acta 1666:40–50
Nakagaki M, Katoh I, Handa T (1981) Surface potential of lipid membrane estimated from the partitioning of methylene blue into liposomes. Biochemistry 20:2208–2212
Fernández-Vidal M, White SH, Ladokhin AS (2011) Membrane partitioning: “classical” and “nonclassical” hydrophobic effects. J Membr Biol 239:5–14
Silvius JR, Nabi IR (2006) Fluorescence-quenching and resonance energy transfer studies of lipid microdomains in model and biological membranes. Mol Membr Biol 23:5–16
Wang T, Cady S, Hong M (2012) NMR determination of protein partitioning into membrane domains with different curvatures and application to the influenza M2 peptide. Biophys J 102:787–794
Scheidt HA, Huster D (2008) The interaction of small molecules with phospholipid membranes studied by 1H NOESY NMR under magic-angle spinning. Acta Pharmacol Sin 29:35–49
Lee TZ, Mozsolits H, Aguilar MI (2001) Measurement of the affinity of melittin for zwitterionic and anionic membranes using immobilised lipid biosensors. J Pept Res 58:464–476
Heerklotz HH, Binder H, Epand RM (1999) A “release” protocol for isothermal titration calorimetry. Biophys J 76:2606–2613
Tsamaloukas A, Keller S, Heerklotz H (2007) Uptake and release protocol for assessing membrane binding and permeation by way of isothermal titration calorimetry. Nat Protoc 2:695–704
Aveyard R, Haydon DA (1973) An introduction to the principles of surface chemistry. Cambridge University Press, Cambridge
McLaughlin S (1977) Electrostatic potentials at membrane-solution interfaces. Curr Top Membr 9:71–144
McLaughlin S (1989) The electrostatic properties of membranes. Annu Rev Biophys Biophys Chem 18:113–136
Keller S, Heerklotz H, Blume A (2006) Monitoring lipid membrane translocation of sodium dodecyl sulfate by isothermal titration calorimetry. J Am Chem Soc 128:1279–1286
Keller S, Böthe M, Bienert M et al (2007) A simple fluorescence-spectroscopic membrane translocation assay. Chembiochem 8:546–552
Bárány-Wallje E, Keller S, Serowy S et al (2005) A critical reassessment of penetratin translocation across lipid membranes. Biophys J 89:2513–2521
Cambridge SB, Geissler D, Keller S et al (2006) A caged doxycycline analogue for photoactivated gene expression. Angew Chem Int Ed 45:2229–2231
Hagen V, Dekowski B, Nache V et al (2005) Coumarinylmethyl esters for ultrafast release of high concentrations of cyclic nucleotides upon one- and two-photon photolysis. Angew Chem Int Ed 44:7887–7891
Gilbert D, Funk K, Lechler R et al (2007) Caged capsaicins: new tools for the examination of TRPV1 channels in somatosensory neurons. Chembiochem 8:89–97
Geissler D, Antonenko YN, Schmidt R et al (2005) (Coumarin-4-yl) methyl esters as highly efficient, ultrafast phototriggers for protons and their application to acidifying membrane surfaces. Angew Chem Int Ed 44:1195–1198
Keller S, Heerklotz H, Jahnke N et al (2006) Thermodynamics of lipid membrane solubilization by sodium dodecyl sulfate. Biophys J 90:4509–4521
Heerklotz H, Tsamaloukas AD, Keller S (2009) Monitoring detergent-mediated solubilization and reconstitution of lipid membranes by isothermal titration calorimetry. Nat Protoc 4:686–697
Krylova OO, Jahnke N, Keller S (2010) Membrane solubilisation and reconstitution by octylglucoside: comparison of synthetic lipid and natural lipid extract by isothermal titration calorimetry. Biophys Chem 150:105–111
Broecker J, Keller S (2010) Membrane translocation assayed by fluorescence spectroscopy. Methods Mol Biol 606:271–289
Kemmer G, Keller S (2010) Nonlinear least-squares data fitting in Excel spreadsheets. Nat Protoc 5:267–281
Keller S, Vargas C, Zhao H et al (2012) High-precision isothermal titration calorimetry with automated peak-shape analysis. Anal Chem 84:5066–5073
Tocanne J-F, Teissie J (1990) Ionization of phospholipids and phospholipid-supported interfacial lateral diffusion of protons in membrane model systems. Biochim Biophys Acta 1031:111–142
Heerklotz H (2001) Membrane stress and permeabilization induced by asymmetric incorporation of compounds. Biophys J 81:184–195
Altenbach C, Seelig J (1984) Ca2+ Binding to phosphatidylcholine bilayers as studied by deuterium magnetic resonance. Evidence for the formation of a Ca2+ complex with two phospholipid molecules. Biochemistry 23:3913–3920
Nagle JF, Tristram-Nagle S (2000) Structure of lipid bilayers. Biochim Biophys Acta 1469:159–195
Acknowledgments
We thank Sebastian Fiedler, Martin Textor, and Sebastian Unger (all University of Kaiserslautern) for helpful discussions and comments. This work was supported by the Phospholipid Research Centre, the Research Initiative Membrane Biology, and the Stiftung Rheinland-Pfalz für Innovation (grant 961-386261/969 to S.K.).
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Vargas, C., Klingler, J., Keller, S. (2013). Membrane Partitioning and Translocation Studied by Isothermal Titration Calorimetry. In: Rapaport, D., Herrmann, J. (eds) Membrane Biogenesis. Methods in Molecular Biology, vol 1033. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-487-6_16
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DOI: https://doi.org/10.1007/978-1-62703-487-6_16
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