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Interakcija egerolizinskega proteina erilizina A z lipidnimi membranami, ki vsebujejo kardiolipin
ID Žeželj, Luka (Author), ID Sepčić, Kristina (Mentor) More about this mentor... This link opens in a new window, ID Resnik, Nataša (Co-mentor)

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Abstract
Protein erilizin A (EryA), izoliran iz gobe Pleurotus eryngii, se močno veže na membrane, ki vsebujejo ceramid fosfoetanolamin in holesterol. Pred kratkim so potrdili vezavo fluorescenčno označenega EryA (EryA-EGFP) na kardiolipin, lipid, ki ga najdemo v bakterijskih membranah, v mitohondrijih ter v kloroplastih. Kardiolipin vpliva na strukturo in delovanje membran ter je udeležen pri uravnavanju programirane celične smrti (apoptoze), saj se v apoptotskih celicah prestavi iz notranje mitohondrijske membrane v zunanjo mitohondrijsko membrano in od tod na plazmalemo. V sklopu magistrske naloge smo z metodo površinske plazmonske resonance analizirali interakcijo EryA z lipidnimi vezikli, ki vsebujejo različne molske deleže kardiolipina. Dokazali smo pozitivno korelacijo med koncentracijo kardiolipina in jakostjo vezave EryA. Prav tako smo spremljali vpliv molskega deleža kardiolipina na urejenost membran, kjer smo s fluorescenčno anizotropijo uspešno dokazali negativni vpliv kardiolipina na ta parameter. V okviru magistrske naloge smo z EryA-mCherry tudi uspešno označili apoptotske epitelijske celice in vitro ter najverjetneje dokazali transport kardiolipina na plazmalemo med apoptozo.

Language:Slovenian
Keywords:kardiolipin, ceramid fosfoetanolamin, sfingomielin, lipidna membrana, erilizin A, apoptoza
Work type:Master's thesis/paper (mb22)
Organization:BF - Biotechnical Faculty
Year:2022
PID:20.500.12556/RUL-139944 This link opens in a new window
Publication date in RUL:09.09.2022
Views:99
Downloads:35
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Secondary language

Language:English
Title:Interaction of the egerolysin protein erylysin A with cardiolipin-containing lipid membranes
Abstract:
The protein erylysin A (EryA), isolated from the mushroom Pleurotus eryngii, binds strongly to membranes containing ceramide phosphoethanolamine and cholesterol. Recently, the binding of fluorescently labeled EryA (EryA-EGFP) to cardiolipin, a lipid found in bacterial membranes, mitochondria and chloroplasts, was confirmed. Cardiolipin affects the structure and function of membranes and is involved in the regulation of programmed cell death (apoptosis), where it is transferred from the inner mitochondrial membrane to the outer mitochondrial membrane and from there to the plasmalemma. We analyzed the interaction of EryA with lipid vesicles containing different mole ratios of cardiolipin using the surface plasmon resonance method. We demonstrated a positive correlation between cardiolipin concentration and EryA binding intensity. We also monitored the influence of the molar ratio of cardiolipin on the membrane ordering, where we successfully demonstrated the negative influence of cardiolipin on this parameter using fluorescence anisotropy. Finally, we successfully labeled apoptotic (mammalian) MDCK cells with EryA-mCherry and most likely proved the transport of cardiolipin to the plasmalemma during apoptosis.

Keywords:cardiolipin, ceramide phosphoethanolamine, sphingomyelin, lipid membrane, erylysin A, apoptosis

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