TY - JOUR
T1 - Spin-label saturation-recovery EPR at W-band
T2 - Applications to eye lens lipid membranes
AU - Mainali, Laxman
AU - Raguz, Marija
AU - Camenisch, Theodore G.
AU - Hyde, James S.
AU - Subczynski, Witold K.
PY - 2011/9
Y1 - 2011/9
N2 - Saturation-recovery (SR) EPR at W-band (94 GHz) to obtain profiles of the membrane fluidity and profiles of the oxygen transport parameter is demonstrated for lens lipid membranes using phosphatidylcholine (n-PC), stearic acid (n-SASL), and cholesterol analog (ASL and CSL) spin labels, and compared with results obtained in parallel experiments at X-band (9.4 GHz). Membranes were derived from the total lipids extracted from 2-year-old porcine lens cortex and nucleus. Two findings are especially significant. First, measurements of the spin-lattice relaxation times T1 for n-PCs allowed T1 profiles across the membrane to be obtained. These profiles reflect local membrane properties differently than profiles of the order parameter. Profiles obtained at W-band are, however, shifted to longer T1 values compared to those obtained at X-band. Second, using cholesterol analog spin labels and relaxation agents (hydrophobic oxygen and water-soluble NiEDDA), the cholesterol bilayer domain was discriminated in membranes made from lipids of the lens nucleus. However, membranes made from cortical lipids show a single homogeneous environment. Profiles of the oxygen transport parameter obtained from W-band measurements are practically identical to those obtained from X-band measurements, and are very similar to those obtained earlier at X-band for membranes made of 2-year-old bovine cortical and nuclear lens lipids (M. Raguz, J. Widomska, J. Dillon, E.R. Gaillard, W.K. Subczynski, Biochim. Biophys. Acta 1788 (2009) 2380-2388). Results demonstrate that SR EPR at W-band has the potential to be a powerful tool for studying samples of small volume, ∼30 nL, compared with the sample volume of ∼3 μL at X-band.
AB - Saturation-recovery (SR) EPR at W-band (94 GHz) to obtain profiles of the membrane fluidity and profiles of the oxygen transport parameter is demonstrated for lens lipid membranes using phosphatidylcholine (n-PC), stearic acid (n-SASL), and cholesterol analog (ASL and CSL) spin labels, and compared with results obtained in parallel experiments at X-band (9.4 GHz). Membranes were derived from the total lipids extracted from 2-year-old porcine lens cortex and nucleus. Two findings are especially significant. First, measurements of the spin-lattice relaxation times T1 for n-PCs allowed T1 profiles across the membrane to be obtained. These profiles reflect local membrane properties differently than profiles of the order parameter. Profiles obtained at W-band are, however, shifted to longer T1 values compared to those obtained at X-band. Second, using cholesterol analog spin labels and relaxation agents (hydrophobic oxygen and water-soluble NiEDDA), the cholesterol bilayer domain was discriminated in membranes made from lipids of the lens nucleus. However, membranes made from cortical lipids show a single homogeneous environment. Profiles of the oxygen transport parameter obtained from W-band measurements are practically identical to those obtained from X-band measurements, and are very similar to those obtained earlier at X-band for membranes made of 2-year-old bovine cortical and nuclear lens lipids (M. Raguz, J. Widomska, J. Dillon, E.R. Gaillard, W.K. Subczynski, Biochim. Biophys. Acta 1788 (2009) 2380-2388). Results demonstrate that SR EPR at W-band has the potential to be a powerful tool for studying samples of small volume, ∼30 nL, compared with the sample volume of ∼3 μL at X-band.
KW - Cholesterol
KW - Cholesterol bilayer domain
KW - EPR
KW - Eye lens
KW - Spin-label
KW - W-band
UR - http://www.scopus.com/inward/record.url?scp=84855404181&partnerID=8YFLogxK
U2 - 10.1016/j.jmr.2011.06.014
DO - 10.1016/j.jmr.2011.06.014
M3 - Article
C2 - 21745756
AN - SCOPUS:84855404181
SN - 1090-7807
VL - 212
SP - 86
EP - 94
JO - Journal of Magnetic Resonance
JF - Journal of Magnetic Resonance
IS - 1
ER -