Optimization of Bicelle Lipid Composition and Temperature for EPR Spectroscopy of Aligned Membranes
Identifieur interne : 000271 ( Main/Exploration ); précédent : 000270; suivant : 000272Optimization of Bicelle Lipid Composition and Temperature for EPR Spectroscopy of Aligned Membranes
Auteurs : Jesse E. Mccaffrey [États-Unis] ; Zachary M. James [États-Unis] ; David D. Thomas [États-Unis]Source :
- Journal of magnetic resonance (San Diego, Calif. : 1997) [ 1090-7807 ] ; 2014.
Abstract
We have optimized the magnetic alignment of phospholipid bilayered micelles (bicelles) for EPR spectroscopy, by varying lipid composition and temperature. Bicelles have been extensively used in NMR spectroscopy for several decades, in order to obtain aligned samples in a near-native membrane environment and take advantage of the intrinsic sensitivity of magnetic resonance to molecular orientation. Recently, bicelles have also seen increasing use in EPR, which offers superior sensitivity and orientational resolution. However, the low magnetic field strength (less than 1 T) of most conventional EPR spectrometers results in homogeneously oriented bicelles only at a temperature well above physiological. To optimize bicelle composition for magnetic alignment at reduced temperature, we prepared bicelles containing varying ratios of saturated (DMPC) and unsaturated (POPC) phospholipids, using EPR spectra of a spin-labeled fatty acid to assess alignment as a function of lipid composition and temperature. Spectral analysis showed that bicelles containing an equimolar mixture of DMPC and POPC homogeneously align at 298 K, 20 K lower than conventional DMPC-only bicelles. It is now be possible to perform EPR studies of membrane protein structure and dynamics in well-aligned bicelles at physiological temperatures and below.
Url:
DOI: 10.1016/j.jmr.2014.09.026
PubMed: 25514061
PubMed Central: 4286475
Affiliations:
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Mccaffrey</name><affiliation wicri:level="2"><nlm:aff id="A1"> Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Minnesota</region></placeName><wicri:cityArea> Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis</wicri:cityArea></affiliation></author><author><name sortKey="James, Zachary M" sort="James, Zachary M" uniqKey="James Z" first="Zachary M." last="James">Zachary M. James</name><affiliation wicri:level="2"><nlm:aff id="A1"> Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Minnesota</region></placeName><wicri:cityArea> Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis</wicri:cityArea></affiliation></author><author><name sortKey="Thomas, David D" sort="Thomas, David D" uniqKey="Thomas D" first="David D." last="Thomas">David D. Thomas</name><affiliation wicri:level="2"><nlm:aff id="A1"> Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455</nlm:aff><country xml:lang="fr">États-Unis</country><placeName><region type="state">Minnesota</region></placeName><wicri:cityArea> Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis</wicri:cityArea></affiliation></author></analytic><series><title level="j">Journal of magnetic resonance (San Diego, Calif. : 1997)</title><idno type="ISSN">1090-7807</idno><idno type="eISSN">1096-0856</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p id="P1">We have optimized the magnetic alignment of phospholipid bilayered micelles (bicelles) for EPR spectroscopy, by varying lipid composition and temperature. Bicelles have been extensively used in NMR spectroscopy for several decades, in order to obtain aligned samples in a near-native membrane environment and take advantage of the intrinsic sensitivity of magnetic resonance to molecular orientation. Recently, bicelles have also seen increasing use in EPR, which offers superior sensitivity and orientational resolution. However, the low magnetic field strength (less than 1 T) of most conventional EPR spectrometers results in homogeneously oriented bicelles only at a temperature well above physiological. To optimize bicelle composition for magnetic alignment at reduced temperature, we prepared bicelles containing varying ratios of saturated (DMPC) and unsaturated (POPC) phospholipids, using EPR spectra of a spin-labeled fatty acid to assess alignment as a function of lipid composition and temperature. Spectral analysis showed that bicelles containing an equimolar mixture of DMPC and POPC homogeneously align at 298 K, 20 K lower than conventional DMPC-only bicelles. It is now be possible to perform EPR studies of membrane protein structure and dynamics in well-aligned bicelles at physiological temperatures and below.</p></div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Minnesota</li></region></list><tree><country name="États-Unis"><region name="Minnesota"><name sortKey="Mccaffrey, Jesse E" sort="Mccaffrey, Jesse E" uniqKey="Mccaffrey J" first="Jesse E." last="Mccaffrey">Jesse E. Mccaffrey</name></region><name sortKey="James, Zachary M" sort="James, Zachary M" uniqKey="James Z" first="Zachary M." last="James">Zachary M. James</name><name sortKey="Thomas, David D" sort="Thomas, David D" uniqKey="Thomas D" first="David D." last="Thomas">David D. Thomas</name></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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