20.500.12556/RUL-177547 Analiza membransko aktivnih proteinov iz gliv za razvoj novih lipidnih označevalcev in biopesticidov Analysis of membrane-active proteins from fungi for the development of new lipid markers and biopesticides Za predstavnike egerolizinske družine proteinov je značilno, da prepoznajo in se vežejo na specifične membranske lipide ali lipidne mešanice, zaradi česar se jih lahko uporablja kot označevalce membranskih lipidov. Čeprav so egerolizini prisotni v vseh domenah življenja, so najbolj raziskani tisti, ki izhajajo iz glivnega rodu Pleurotus. Večina organizmov z zapisi za egerolizine, vsebuje tudi zapise za partnerske proteine z domeno kompleksa, ki napada membrane/perforin (MACPF). Mešanica egerolizina in partnerskega proteina je litična za celice, ki v svojih membranah vsebujejo lipidni receptor za egerolizin, zaradi česar jih je mogoče uporabiti kot biopesticide. V doktorski nalogi smo rekombinantno izrazili osem novih egerolizinov in pet proteinov z domeno MACPF iz bazidiomicet Pleurotus pulmonarius, Heterobasidion irregulare, Trametes versicolor, Mucidula mucida in Lepista nuda, ter jih primerjali z že raziskanim egerolizinom ostreolizinom A6 in njegovim proteinskim partnerjem pleurotolizinom B iz bazidiomicetne glive bukovega ostrigarja (Pleurotus ostreatus). Večina izmed teh novo opisanih egerolizinov tvori v kombinaciji s svojimi proteinskimi partnerji citolitične komplekse v membranah, ki vsebujejo specifične sfingolipidne receptorje. Nekateri egerolizini so hkrati sposobni vezave na nekatere glicerofosfolipide, zlasti na fosfatidno kislino in kardiolipin, vendar te vrste interakcij ne vodijo do nastanka por. Rezultati raziskave nakazujejo, da bi bili nekateri izmed na novo opisanih egerolizinov lahko novi molekularni biooznačevalci lipidov, v kombinaciji s proteinskimi partnerji z domeno MACPF pa bi bili nekateri egerolizini lahko uporabn tudi kot novi biopesticidi. Aegerolysins are a family of proteins that recognize and bind to specific membrane lipids or lipid mixtures, making them suitable as membrane lipid markers. Although they are present across all domains of life, those produced by fungi of the genus Pleurotus are the most well-studied. Most of these fungi harbour also the genes encoding proteins that contain the membrane attack complex/perforin (MACPF) domain. The combination of aegerolysin and its MACPF protein partner is lytic to cells that possess the aegerolysin lipid receptor, which makes them potential biopesticides. In the doctoral thesis, we recombinantly expressed eight novel aegerolysin proteins and five MACPF-domain proteins from the basidiomycetes Pleurotus pulmonarius, Heterobasidion irregulare, Trametes versicolor, Mucidula mucida, and Lepista nuda, and compared the activities of these proteins with the most studied aegerolysin, ostreolysin A6 and its MACPF protein partner pleurotolysin B from P. ostreatus. Most of these new proteins form active aegerolysin/MACPF cytolytic complexes upon binding of aegerolysin to specific sphingolipid-containing membranes. Some aegerolysins can also bind to selected glycerophospholipids, particularly phosphatidic acid and cardiolipin, but these interactions do not lead to pore formation. Our results indicate that some of these selected aegerolysins show potential as new molecular biosensors of membrane lipids or as biopesticides. egerolizini MACPF proteini membrane lipidi glive pore citolitični kompleksi aegerolysins MACPF proteins membranes lipids fungi pores cytolytic complexes true false false [L. L. Popošek] Slovenski jezik Angleški jezik Doktorsko delo/naloga 2025-12-24 07:16:14 2025-12-24 07:16:20 2026-01-06 11:54:43 0000-00-00 00:00:00 2025 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 577.126(043.3) 277658 263565571 55614.pdf 55614.pdf 1 832504394D77E94E4C48647EE742AB1B eaf7fcfa0e1a56096366c4650e2c46a9962fb575c995e57f65d9b42f8b0693d1 9bbf63e0-e08f-11f0-9328-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=224619 Biotehniška fakulteta 0 0 0