20.500.12556/RUL-180918 Sensing of cardiolipin exposure on plasma membranes of apoptotic cells by EryA-mCherry protein Erylysin A (EryA), an aegerolysin protein produced by the edible king oyster mushroom (Pleurotus eryngii), interacts strongly with an invertebrate-specific membrane sphingolipid ceramide phosphoethanolamine. Recently, a fluorescently fused variant of EryA was shown to bind to artificial and bacterial lipid membranes containing cardiolipin (CL). This tetra-acylated glycerophospholipid, present in bacteria and in inner mitochondrial mem-branes of eukaryotic cells, was shown to be externalized to the plasma membrane surface during the process of apoptosis. In this work, we evaluated the interaction of EryA-mCherry with CL-containing artificial lipid vesicles and with mammalian cells undergoing apoptosis and compared its binding affinity and specificity to that of the well-established apoptosis marker, annexin V-FITC. Our results show that, in contrast to annexin V-FITC, which binds several negatively charged glycerophospholipids, EryA-mCherry specifically recognizes and binds CL in artificial membrane systems. However, this binding of EryA-mCherry to CL-supplemented membranes is less effective (K$_D$ = 4.7 ±  1.6 μM) than that of annexin V-FITC, whose binding is observed at nanomolar concentrations. Experiments using mammalian cells showed the ability of EryA-mCherry to selectively label the membranes of apoptotic cells, binding to the same membrane regions as anti-CL antibodies and annexin V-FITC. Our data suggest that EryA-mCherry might be used as a marker of early apoptosis, as well as a marker of CL in biological and artificial lipid membranes. aegerolysin annexin V apoptosis marker cardiolipin Pleurotus lipidne membrane kardiolipin egerolizini biološki označevalci true false true Angleški jezik Slovenski jezik Članek v reviji 2026-03-19 12:33:41 2026-03-19 12:33:48 2026-03-22 04:08:38 0000-00-00 00:00:00 2026 0 0 Str. 1150-1167 iss. 4 Vol. 293 Feb. 2026 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577.112:577.29:601.2 1742-4658 10.1111/febs.70290 254650115 The_FEBS_Journal_-_2025_-_Zezelj_-_Sensing_of_cardiolipin_exposure_on_plasma_membranes_of_apoptotic_cells_by_EryA‐mCherry.pdf The_FEBS_Journal_-_2025_-_Zezelj_-_Sensing_of_cardiolipin_exposure_on_plasma_membranes_of_apoptotic_cells_by_EryA‐mCherry.pdf 1 68A92372F73BE92C9B8B4367702ACCFB 1ef74c803313392ae44ef913fb9b42fe051aa67ad209a72df7ef60310ffa1c94 e69c332b-2387-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=230891 https://febs.onlinelibrary.wiley.com/doi/10.1111/febs.70290 1 69caceb2-2387-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=230888 Biotehniška fakulteta Medicinska fakulteta 0 0 0