Saturation-recovery electron paramagnetic resonance discrimination by oxygen transport (DOT) method for characterizing membrane domains

Methods in Molecular Biology
Witold K SubczynskiAkihiro Kusumi

Abstract

The discrimination by oxygen transport (DOT) method is a dual-probe saturation-recovery electron paramagnetic resonance approach in which the observable parameter is the spin-lattice relaxation time (T1) of lipid spin labels, and the measured value is the bimolecular collision rate between molecular oxygen and the nitroxide moiety of spin labels. This method has proven to be extremely sensitive to changes in the local oxygen diffusion-concentration product (around the nitroxide moiety) because of the long T1 of lipid spin labels (1-10 micros) and also because molecular oxygen is a unique probe molecule. Molecular oxygen is paramagnetic, small, and has the appropriate level of hydrophobicity that allows it to partition into various supramolecular structures such as different membrane domains. When located in two different membrane domains, the spin label alone most often cannot differentiate between these domains, giving very similar (indistinguishable) conventional electron paramagnetic resonance spectra and similar T1 values. However, even small differences in lipid packing in these domains will affect oxygen partitioning and oxygen diffusion, which can be easily detected by observing the different T1s from spin labels in thes...Continue Reading

Citations

Dec 31, 2011·The Journal of Membrane Biology·Witold K SubczynskiAlexey Konovalov
Jul 1, 2017·Cell Biochemistry and Biophysics·Justyna WidomskaMarija Raguz
May 29, 2018·Applied Magnetic Resonance·Laxman MainaliWitold K Subczynski
Apr 25, 2007·Biochimica Et Biophysica Acta·Justyna WidomskaWitold K Subczynski
Feb 27, 2008·Biochimica Et Biophysica Acta·Marija RaguzWitold K Subczynski
Jul 13, 2011·Journal of Magnetic Resonance·Laxman MainaliWitold K Subczynski

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