20.500.12556/RUL-156325 Sequestration of membrane cholesterol by cholesterol-binding proteins inhibits SARS-CoV-2 entry into Vero E6 cells Membrane lipids and proteins form dynamic domains crucial for physiological and pathophysiological processes, including viral infection. Many plasma membrane proteins, residing within membrane domains enriched with cholesterol (CHOL) and sphingomyelin (SM), serve as receptors for attachment and entry of viruses into the host cell. Among these, human coronaviruses, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), use proteins associated with membrane domains for initial binding and internalization. We hypothesized that the interaction of lipid-binding proteins with CHOL in plasma membrane could sequestrate lipids and thus affect the efficiency of virus entry into host cells, preventing the initial steps of viral infection. We have prepared CHOL-binding proteins with high affinities for lipids in the plasma membrane of mammalian cells. Binding of the perfringolysin O domain four (D4) and its variant D4$^{E458L}$ to membrane CHOL impaired the internalization of the receptor-binding domain of the SARS-CoV-2 spike protein and the pseudovirus complemented with the SARS-CoV-2 spike protein. SARS-CoV-2 replication in Vero E6 cells was also decreased. Overall, our results demonstrate that the integrity of CHOL-rich membrane domains and the accessibility of CHOL in the membrane play an essential role in SARS-CoV-2 cell entry. cholesterol endocytosis plasma membrane membrane domains pore-forming proteins SARS-CoV-2 viral infection biokemija SARS-CoV-2 holesterol endocitoza beljakovine koronavirusi true false true Angleški jezik Slovenski jezik Članek v reviji 2024-05-20 11:35:00 2024-05-20 11:35:02 2024-05-21 03:46:45 0000-00-00 00:00:00 2024 0 0 16 str. Vol. 716, art. 149954 2024 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 1090-2104 10.1016/j.bbrc.2024.149954 194174467 1-s2.0-S0006291X2400490X-main.pdf 1-s2.0-S0006291X2400490X-main.pdf 1 3E488C33DAFB10874324928F09DE89CA fa1615e42cdaf446be460e726109fdfd979f7214df20fc867ed61879296d4218 981a330a-168c-11ef-b600-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=183725 https://www.sciencedirect.com/science/article/pii/S0006291X2400490X 1 3b098954-168c-11ef-b600-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=183724 Veterinarska fakulteta 0 0 0