Interakcije egerolizinskih proteinov iz gliv rodu Pleurotus z lipidi iz bakterij, kvasovk in nitastih gliv

Zidar, Luen (avtor), Sepčić, Kristina (mentor), Skočaj, Matej (komentor)

Ostreolizin A (OlyA) iz gobe bukov ostrigar (Pleurotus ostreatus) uvrščamo v proteinsko družino egerolizinov. OlyA prepozna in se veže na membranske domene obogatene s holesterolom in sfingomielinom, t.i. lipidne rafte ter na ceramid fosfoetanolamin (CPE), sfingolipid, ki je specifičen za nevretenčarje in nekatere po Gramu negativne bakterije. Zaradi omenjenih lastnosti imajo OlyA in drugi egerolizini velik biotehnološki potencial, ki se kaže tudi v klinični diagnostiki in v kmetijstvu za zatiranje škodljivcev, saj lahko nekateri egerolizini v kombinaciji s partnerskim proteinom delujejo kot tvorci por s hemolitičnim in citolitičnim delovanjem. Za uspešno uporabo egerolizinov in razvijanje potencialnih aplikacij, je ključnega pomena poznavanje njihovih tarč in specifik vezave. Z metodo točkovnega nanosa smo preverili interakcijo fluorescenčno označenega egerolizina (OlyA-mCherry) s komponentami biofilmov različnih bakterijskih sevov in ugotovili, da v biofilmu teh sevov ni tarč, ki bi jih OlyA prepoznal in na katere bi se vezal. Z metodo točkovnega nanosa in metodo prenosa far-eastern smo preverili interakcijo OlyA-mCherry s polarnimi in nepolarnimi lipidi iz bakterij, kvasovk, nitastih gliv in ene vrste arhej. Očitno interakcijo smo dokazali le z nepolarnimi lipidi bakterije Prevotella sp. Signal je bil zaznan tudi v primeru interakcije OlyA s polarno in nepolarno frakcijo lipidov iz P. aeruginosa EXB L-1125, vendar so ti rezultati nezanesljivi. Na podlagi rezultatov lahko z veliko verjetnostjo ponovno potrdimo specifičnost vezave OlyA s CPE in odsotnost dodatnih membranskih tarč pri veliki večini testiranih mikroorganizmov. Pokazali smo, da OlyA ne interagira s komponentami bakterijskega biofilma in prav tako ne s polarnimi oz. nepolarnimi lipidi iz testiranih bakterij, kvasovk, nitastih gliv in ene vrste arhej. Dobljeni rezultati omogočajo nadaljevanje razvoja uporabe egerolizinov v različnih biotehnoloških aplikacijah.

• Jezik: Slovenski jezik
• Ključne besede: egerolizini, ostreolizin A, OlyA-mCherry, lipidi, membranske tarče, ceramid fosfoetanolamin, bakterijski biofilm
• Vrsta gradiva: Magistrsko delo/naloga
• Organizacija: BF - Biotehniška fakulteta
• Leto izida: 2021
• PID: 20.500.12556/RUL-124706
• COBISS.SI-ID: 52364547
• Datum objave v RUL: 11.02.2021

ZIDAR, Luen, 2021, Interakcije egerolizinskih proteinov iz gliv rodu Pleurotus z lipidi iz bakterij, kvasovk in nitastih gliv [na spletu]. Magistrsko delo. [Dostopano 11 april 2025]. Pridobljeno s: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=slv&id=124706

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Interactions of aegerolysin proteins from fungi of the genus Pleurotus with lipids from bacteria, yeasts and filaments

Izvleček

Ostreolysin A (OlyA) from oyster mushroom (Pleurotus ostreatus) is a member of the aegerolysin protein family. OlyA recognizes and binds to membrane domains enriched in cholesterol and sphingomyelin, so-called lipid rafts, and to ceramide phosphoethanolamine (CPE), a sphingolipid specific for invertebrates and some Gram negative bacteria. Due to previously mentioned features, OlyA and other aegerolysins have great biotechnological potential which is particulary shown in fields of clinical diagnostics and agriculture. Some of the aegerolysins, combined with their partner proteins, can act as pore formers and cam exert hemolytic and cytolytic effects. The key to successful use of aegerolysins and development of potential applications is in identifying their targets and in understanding their binding specificity. Using the dot blot method, we tested the interaction between fluorescently-labelled aegerolysin (OlyA-mCherry) and biofilm components of different strains of bacteria. We discovered bacterial biofilms do not contain targets that OlyA would recognize and bind. Using the dot blot and the far-eastern blot method, we examined the interaction of OlyA-mCherry with polar and non-polar lipids in bacteria, yeasts, filamentous fungus and in one species of archaea. The only apparent interaction was observed with non-polar lipids of a bacterium Prevotella spp. A signal was also shown in the case of OlyA interaction with polar and non-polar lipid fraction in P. aeruginosa EXB L-1125; however, these results are not reliable. According to the results we can, with great probability, confirm the specificity of OlyA interaction with CPE and the absence of additonal membrane targets in the majority of the tested microorganisms. We demonstrated that OlyA does not interact with bacterial biofilm components, nor with polar and non-polar lipids in bacteria, yeasts, filamentous fungus and archaea. The results thus allow further development of the use of aegerolysins in different biotechnological applications.

• Ključne besede: aegerolysins, ostreolysine A, OlyA-mCherry, lipids, membrane targets, ceramide phosphoethanolamine, bacterial biofilms