Analiza membransko aktivnih proteinov iz gliv za razvoj novih lipidnih označevalcev in biopesticidov : raziskovalni podatki, obravnavani v doktorskem delu

Popošek, Larisa Lara (author)

V raziskavi smo biokemijsko opredelili egerolizine in njihove proteinske partnerje z domeno MACPF iz izbranih bazidiomicet, pri čemer je bil glavni poudarek na uporabljenih metodah. Najprej smo izvedli bioinformatsko analizo za določitev osnovnih biokemijskih lastnosti proteinov, nato pa pripravili rekombinantne proteine za eksperimentalno delo. Strukturne lastnosti smo preučevali s CD-spektrometrijo. Za opredelitev interakcij z membranami smo uporabili pripravo lipidnih mešanic, sedimentacijski test in površinsko plazmonsko resonanco (SPR). Funkcionalno aktivnost citolitičnih kompleksov smo analizirali s testom sproščanja kalceina, testom hemolize ter z določitvijo citotoksičnosti na celični liniji Sf9. Na koncu smo izvedli še teste toksičnosti na žuželkah. S temi metodami smo ovrednotili strukturne, vezavne in funkcionalne lastnosti preučevanih proteinov.

• Language: Slovenian
• Keywords: egerolizini, MACPF proteini, membrane, lipidi, citolitični kompleksi
• Typology: 2.20 - Complete scientific database of research data
• Geographic coverage: Slovenija
• Time coverage: Od/From 1. 10. 2021 do/till 30. 9. 2025
• Organization: BF - Biotechnical Faculty
• Year: 2026
• PID: 20.500.12556/RUL-181284
• Data col. methods: Experiment; Experiment: Laboratory; Experiment: Field/Intervention; Measurements and tests
• Publication date in RUL: 07.04.2026

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: English
• Title: Analysis of membrane-active proteins from fungi for the development of new lipid markers and biopesticides : research data underlying the doctoral dissertation

Abstract

In this study, we biochemically characterized egerolysins and their protein partners containing the MACPF domain from selected basidiomycetes, with a primary focus on the methods used. First, we performed a bioinformatic analysis to determine the basic biochemical properties of the proteins, followed by the preparation of recombinant proteins for experimental work. Structural properties were analyzed using CD spectroscopy. To investigate membrane interactions, we used lipid mixture preparation, sedimentation assays, and surface plasmon resonance (SPR). The functional activity of cytolytic complexes was assessed using a calcein release assay, a hemolysis assay, and cytotoxicity testing on the Sf9 cell line. Finally, toxicity assays were performed on insects. Using these methods, we evaluated the structural, binding, and functional properties of the studied proteins.

• Keywords: aegerolysins, MACPF proteins, membranes, lipids, cytolytic complexes

Projects

• Funder: ARIS - Slovenian Research and Innovation Agency
• Project number: P1-0207
• Name: Toksini in biomembrane