Dissecting lipid contents in the distinct regions of native retinal rod disk membranes

Sander, C. L.; Sears, A. E.; Pinto, A. F. M.; Choi, E. H.; Kahremany, S. L.; Jin, H. G.; Suh, S.; Pardon, E.; Dong, Z.; Steyaert, J.; Saghatelian, A.; Skowronska-Krawczyk, D.; Kiser, P. D.; Palczewski, K.

doi:10.1101/2021.01.11.426250

bioRxiv · 10.1101/2021.01.11.426250

Dissecting lipid contents in the distinct regions of native retinal rod disk membranes

Sander, C. L.·Sears, A. E.·Pinto, A. F. M.·Choi, E. H.·Kahremany, S. L.·Jin, H. G.·Suh, S.·Pardon, E.·Dong, Z.·Steyaert, J.·Saghatelian, A.·Skowronska-Krawczyk, D.·Kiser, P. D.·Palczewski, K.

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Abstract

Photoreceptors rely on distinct membrane compartments to support their specialized function. Unlike protein localization, identification of critical differences in membrane content has not yet been expanded to lipids, due to the difficulty of isolating domain-specific samples. We have overcome this by using SMA to co-immunopurify membrane proteins and their native lipids from two regions of photoreceptor ROS disks. Each samples copurified lipids were subjected to untargeted lipidomic and fatty acid analysis. Extensive differences between center (rhodopsin) and rim (ABCA4 and PRPH2/ROM1) samples included a lower PC to PE ratio and increased LC- and VLC-PUFAs in the center relative to the rim region, which were enriched in shorter, saturated FAs. The comparatively few differences between the two rim samples likely reflect specific protein-lipid interactions. High-resolution profiling of the ROS disk lipid composition provides a model for future studies of other complex cellular structures, and gives new insights into how intricate membrane structure and protein activity are balanced within the ROS. SUMMARYSander et al. have parsed the lipid composition of native-source photoreceptor disks and find large differences in fatty acid unsaturation and chain length between the center and rim regions. They selectively copurify membrane proteins and lipids from each region in SMALPs using nanobodies and antibodies.

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Sander, C. L., Sears, A. E., Pinto, A. F. M., Choi, E. H., Kahremany, S. L., Jin, H. G., Suh, S., Pardon, E., Dong, Z., Steyaert, J., Saghatelian, A., Skowronska-Krawczyk, D., Kiser, P. D., Palczewski, K.. 2021-01-11. Dissecting lipid contents in the distinct regions of native retinal rod disk membranes. https://doi.org/10.1101/2021.01.11.426250

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