Lipid nanoparticles with erythrocyte cell-membrane proteins

• DOI: 10.1016/j.molliq.2022.120791
• Publication Date: 2022-11-13T22:40:11Z
• Authors: Attila Bóta; Bence Fehér; András Wacha; Tünde Juhász; Dániel Szabó; Lilla Turiák; Anikó Gaál; Zoltán Varga; Heinz Amenitsch; Judith Mihály
• Keywords: 0301 basic medicine; 03 medical and health sciences; QD Chemistry / kémia; Secondary structure; Nanoparticles; Protein mass-pattern; Protein-lipid assembly; Nanoerythrosomes; Protein-to-lipid ratio

When the separated erythrocyte membranes (known as ghosts) are ultrasonicated, a significant part of the membrane proteins are released in the aqueous solvent, instead of being incorporated into the membranes of the formed nanoerythrosomes. In contrast to their membrane-bound counterparts, where helices and β-strands dominate, the released proteins show perturbed secondary structures with an increased ratio of helices, presumably participating in molten globules, as it has been revealed by circular dichroism (CD) and infra-red spectroscopy (IR). The shape and size of these proteins is diverse, and even their aggregates appear. When excess lipid (palmitoyl-lysophosphatidylcholine, LPC) is added to different ghost-derivatives (the full nanoerythrosome system, and its ultracentrifugation pellet and supernatant) in 2 × and 5 × lipid-to-protein mass ratio, various lipid nanoparticles are produced. The core–shell and nanodisc structural models obtained by small-angle X-ray scattering (SAXS) indicate that the choice of the precursor system has a more prominent effect on the resulting shape than the amount of lipid added: when starting from the protein-rich supernatant fraction, small (approx. 5 nm high and 7 nm wide) nanodisks are created. When lipid membranes are already present (in the pellet and the full nanoerythrosome fraction), similar LPC addition results in prolate ellipsoidal particles, with an aspect ratio between 3 and 5, and decreasing overall size when the amount of added lipid is increased. The ellipsoids formed from the total nanoerythrosome fraction are smaller than those from the ultracentrifugation pellet (longest axis around 15 vs 26 nm), whereas for higher LPC-to-protein ratio, the size in both cases reduce to nearly the same (13–14 nm) in both cases.