Membrane Localization and Lipid Interactions of Common Lipid-Conjugated Fluorescence Probes

Oskar Engberg; Holger A Scheidt; Thomas K M Nyholm; J Peter Slotte; Daniel Huster

doi:10.1021/acs.langmuir.9b01202

Membrane Localization and Lipid Interactions of Common Lipid-Conjugated Fluorescence Probes

Langmuir. 2019 Sep 10;35(36):11902-11911. doi: 10.1021/acs.langmuir.9b01202. Epub 2019 Aug 29.

Authors

Oskar Engberg^{1

2}, Holger A Scheidt¹, Thomas K M Nyholm², J Peter Slotte², Daniel Huster¹

Affiliations

¹ Institute for Medical Physics and Biophysics, Medical Department , Leipzig University , Leipzig , Germany.
² Biochemistry, Faculty of Science and Engineering , Åbo Akademi University , Turku , Finland.

PMID: 31424941
DOI: 10.1021/acs.langmuir.9b01202

Abstract

Lateral segregation of lipids in model and biological membranes has been studied intensively in the last decades using a comprehensive set of experimental techniques. Most methods require a probe to report on the biophysical properties of a specific molecule in the lipid bilayer. Because such probes can adversely affect the results of the measurement and perturb the local membrane structure and dynamics, a detailed understanding of probe behavior and its influence on the properties of its direct environment is important. Membrane phase-selective and lipid-mimicking molecules represent common types of probes. Here, we have studied how the fluorescent probes trans-parinaric acid (tPA), diphenylhexatriene (DPH), and 1-oleoyl-2-propionyl[DPH]-sn-glycero-3-phosphocholine (O-DPH-PC) affect the membrane properties of 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine (POPC) bilayers using ²H and ³¹P NMR spectroscopy in the solid state. In addition, using 2D ¹H magic-angle spinning (MAS) nuclear Overhauser enhancement spectroscopy (NOESY) NMR, we have determined the distribution of the probe moieties in the POPC membrane parallel to the membrane normal. We found that the different probes exhibit distinct membrane localizations and distributions, e.g. tPA is located parallel to the membrane normal while DPH predominantly exist in two orientations. Further, tPA was conjugated to sphingomyelin (tPA-SM) as a substitute for the acyl chain in the SM. ¹H NOESY NMR was used to probe the interaction of the tPA-SM with cholesterol as dominant in liquid ordered membrane domains in comparison to POPC-cholesterol interaction in membranes composed of ternary lipid mixtures. We could show that tPA-SM exhibited a strong favorable and very temperature-dependent interaction with cholesterol in comparison to POPC. In conclusion, the NMR techniques can explain probe behavior but also be used to measure lipid-specific affinities between different lipid segments and individual molecules in complex bilayers, relevant to understanding nanodomain formation in biological membranes.

Publication types

Research Support, Non-U.S. Gov't