Lipid mobility and order in bovine rod outer segment disk membranes. A spin-label study of lipid-protein interactions

Abstract

Lipid-protein interactions in bovine rod outer segment disk membranes have been studied by using a series of eight stearic acid spin-label probes which were labeled at different carbon atom positions in the chain. In randomly oriented membrane dispersions, the electron spin resonance (ESR) spectra of the C-8, C-9, C-10, C-11, C-12, C-13, and C-14 atom positional isomers all apparently consist of two components. One of the components corresponds closely to the spectra obtained from dispersions of the extracted membrane lipids, and the other, which is characterized by a considerably greater degree of motional restriction of the lipid chains, is induced by the presence of the protein. Digital subtraction has been used to separate the two components. The proportion of the motionally restricted lipid component is approximately constant, independent of the position of the spin-label group, and corresponds to 30-40% of the total spin-label spectral intensity. The hyperfine splitting of the outer maxima in the difference spectra of the motionally restricted component decreases, and concomitantly, the line widths increase with increasing temperature but change relatively little with increasing distance of the spin-label group from the polar head-group region. This indicates that the corresponding chain motions of the protein-interacting lipids lie in the slow-motion regime of spin-label ESR spectroscopy (tau R approximately 10(-8) S) and that the mobility of these lipids increases with increasing temperature but does not vary greatly along the length of the chain. The data from the hyperfine splittings also suggest the existence of a polarity gradient immediately adjacent to the protein surface, as observed in the fluid lipid regions of the membrane. The more fluid lipid component is only slightly perturbed relative to the lipids alone (for label positions 5-14, inclusive), indicating the presence of chain motions on the nanosecond time scale, and the spectra also reveal a similar polarity profile in both lipid and membrane environments. ESR spectra have also been obtained as a function of magnetic field orientation with oriented membrane samples. For the C-14 atom positional isomer, the motionally restricted component is observed to have a large hyperfine splitting, with the magnetic field oriented both parallel and perpendicular to the membrane normal. This indicates that the motionally restricted lipid chains have a broad distribution of orientations at this label position.(ABSTRACT TRUNCATED AT 400 WORDS)

Integral membrane proteins significantly decrease the molecular motion in lipid bilayers: a deuteron NMR relaxation study of membranes containing myelin proteolipid apoprotein.

“…4 (19,20), in qualitative agreement with the present results. In contrast, fluorescent-probe measurements on the other systems have been interpreted to suggest that an increase in order of the lipid chains is induced by the protein (21,22), contrary to the present direct measurements.…”

Lipid interactions with transmembrane proteins

“…Information about chain orientation and ordering can be obtained from ESR measurements with macroscopically aligned samples [12]. This has been done for oriented rod outer segment discs: it was found that there is a wide orientational distribution, i. e. a considerable degree of disorder, of the lipid chain segments associated with the visual receptor rhodopsin [13].…”

Water Penetration Profile at the Protein-Lipid Interface in Na,K-ATPase Membranes

“…A variety of electron paramagnetic resonance (EPR) parameters are available to characterize the environmental polarity. These include hyperfine couplings in conventional EPR (3)(4)(5), g-values in high-field EPR (6), relaxation enhancement by molecular oxygen (7)(8)(9), and direct detection of penetrant water by pulsed EPR (10,11).…”