“…These materials were obtained by sucrose gradient ultracentrifugation of the Jurkat membrane lysates followed by centrifugal concentration of the top fractions (see Experimental procedures below). As demonstrated in Figures 6 and 7 , the lipid composition of the materials originating from the FFD8-treated membranes was similar to that of SMA-treated membranes and corresponded to the results of our previous study ( 15 ); both were characterized by high cholesterol content and predominance of lipids with saturated fatty acid residues. Figure 6 Comparison of lipid composition of the membrane fragments resistant to SMA versus FFD8.…”
Section: Resultssupporting
confidence: 86%
“…As mentioned above, in our previous studies, we used density gradient ultracentrifugation to demonstrate that the copolymer SMA efficiently solubilizes, presumably into nanodiscs, most membrane proteins except for those present in membrane raft microdomains, which remain in larger and buoyant SMA-resistant membrane fragments ( 7 , 15 ). As shown in Figure 2 , very similar results were obtained with Jurkat cell membranes disintegrated by 1% FFD8—the membrane raft markers (LCK and CD59) were also largely present in peptide-resistant membrane fragments (PRMs) in the top fractions of the gradient.…”
Section: Resultsmentioning
confidence: 98%
“…( 14 )). In our recent studies, we used SMA to characterize membrane proteins present within and outside of membrane rafts in T cells and other cell types ( 7 , 15 ). We also demonstrated similar membrane-disintegrating properties of other amphiphilic copolymers ( 8 , 16 ).…”
“…These materials were obtained by sucrose gradient ultracentrifugation of the Jurkat membrane lysates followed by centrifugal concentration of the top fractions (see Experimental procedures below). As demonstrated in Figures 6 and 7 , the lipid composition of the materials originating from the FFD8-treated membranes was similar to that of SMA-treated membranes and corresponded to the results of our previous study ( 15 ); both were characterized by high cholesterol content and predominance of lipids with saturated fatty acid residues. Figure 6 Comparison of lipid composition of the membrane fragments resistant to SMA versus FFD8.…”
Section: Resultssupporting
confidence: 86%
“…As mentioned above, in our previous studies, we used density gradient ultracentrifugation to demonstrate that the copolymer SMA efficiently solubilizes, presumably into nanodiscs, most membrane proteins except for those present in membrane raft microdomains, which remain in larger and buoyant SMA-resistant membrane fragments ( 7 , 15 ). As shown in Figure 2 , very similar results were obtained with Jurkat cell membranes disintegrated by 1% FFD8—the membrane raft markers (LCK and CD59) were also largely present in peptide-resistant membrane fragments (PRMs) in the top fractions of the gradient.…”
Section: Resultsmentioning
confidence: 98%
“…( 14 )). In our recent studies, we used SMA to characterize membrane proteins present within and outside of membrane rafts in T cells and other cell types ( 7 , 15 ). We also demonstrated similar membrane-disintegrating properties of other amphiphilic copolymers ( 8 , 16 ).…”
“…The ability to observe different types of water molecules in membranes and the feasibility of differentiation of the environment in ion channels by 17 O NMR experiments are unique and exciting. The use of nanodiscs to measure residual 17 O quadrupole couplings to study the structure and dynamics of membrane-bound water and to study the phase transition of encased lipids opens new avenues in membrane biophysics. − The fact that water plays a crucial role in biology and the advent of higher magnetic fields and better NMR instrumentations renders more sophisticated NMR techniques, the benefits of 17 O based NMR applications can be expected to grow remarkably. Therefore, we conclude that much remains to be done to fully utilize the power of 17 O based NMR spectroscopy to study biological solids.…”
Despite the limitations posed by poor sensitivity, studies have reported the unique advantages of 17 O based NMR spectroscopy to study systems existing in liquid, solid, or semisolid states. 17 O NMR studies have exploited the remarkable sensitivity of quadrupole coupling and chemical shift anisotropy tensors to the local environment in the characterization of a variety of intraand intermolecular interactions and motion. Recent studies have considerably expanded the use of 17 O NMR to study dynamic intermolecular interactions associated with some of the challenging biological systems under magic angle spinning (MAS) and aligned conditions. The very fast relaxing nature of 17 O has been well utilized in cellular and in vivo MRS (magnetic resonance spectroscopy) and MRI (magnetic resonance imaging) applications. The main focus of this Review is to highlight the new developments in the biological solids with a detailed discussion for a few selected examples including membrane proteins and nanodiscs. In addition to the unique benefits and limitations, the remaining challenges to overcome, and the impacts of higher magnetic fields and sensitivity enhancement techniques are discussed.
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