20.500.12556/RUL-135717 Dissecting out the molecular mechanism of insecticidal activity of ostreolysin A6/pleurotolysin B complexes on western corn rootworm Ostreolysin A6 (OlyA6) is a protein produced by the oyster mushroom (Pleurotus ostreatus). It binds to membrane sphingomyelin/cholesterol domains, and together with its protein partner, pleurotolysin B (PlyB), it forms 13-meric transmembrane pore complexes. Further, OlyA6 binds 1000 times more strongly to the insect-specific membrane sphingolipid, ceramide phosphoethanolamine (CPE). In concert with PlyB, OlyA6 has potent and selective insecticidal activity against the western corn rootworm. We analysed the histological alterations of the midgut wall columnar epithelium of western corn rootworm larvae fed with OlyA6/PlyB, which showed vacuolisation of the cell cytoplasm, swelling of the apical cell surface into the gut lumen, and delamination of the basal lamina underlying the epithelium. Additionally, cryo-electron microscopy was used to explore the membrane interactions of the OlyA6/PlyB complex using lipid vesicles composed of artificial lipids containing CPE, and western corn rootworm brush border membrane vesicles. Multimeric transmembrane pores were formed in both vesicle preparations, similar to those described for sphingomyelin/cholesterol membranes. These results strongly suggest that the molecular mechanism of insecticidal action of OlyA6/PlyB arises from specific interactions of OlyA6 with CPE, and the consequent formation of transmembrane pores in the insect midgut. aegerolysin bioinsecticide MACPF-protein oyster mushroom pore-forming protein western corn rootworm egerolizin bioinsekticid protein z domeno MACPF bukov ostrigar porotvorni protein koruzni hrošč true false true Angleški jezik Slovenski jezik Članek v reviji 2022-03-29 13:18:16 2022-03-29 13:18:18 2022-09-14 04:14:49 0000-00-00 00:00:00 2021 0 0 16 str. iss. 7, art. 455 Vol. 13 2021 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 577 2072-6651 10.3390/toxins13070455 68691203 RAZ_Milijas_Jotic_Matej_2021.pdf RAZ_Milijas_Jotic_Matej_2021.pdf 1 66572F827B63278602ACF5B688E3D0DA a5d8109d6cdfc84660d417edb36395fe8b6c554582fb0d177b0ce6c7bca6a01e 94d777fb-af52-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=154972 https://www.mdpi.com/2072-6651/13/7/455 1 eb43c05e-af51-11ec-8aca-00155dcfd717 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=154971 Biotehniška fakulteta 0 0 0