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Egerolizini kot molekularno orodje za odkrivanje ceramid fosfoetanolamina, biološkega označevalca s parodontalno boleznijo povezanih bakterij
ID Munjaković, Haris (Author), ID Skočaj, Matej (Mentor) More about this mentor... This link opens in a new window, ID Gašperšič, Rok (Co-mentor)

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Abstract
Egerolizini so majhni (~13-20 kDa), β-strukturni proteini, katerih skupna lastnost je ta, da prepoznajo različne lipide oz. lipidne domene, ki gradijo biološke membrane. Za nekatere glivne egerolizine je znano, da se specifično in močno vežejo na ceramid fosfoetanolamin (CPE). Gre za lipid iz skupine sfingolipidov, ki predstavlja karakteristični lipid v celičnih membranah po Gramu negativnih bakterijskih vrst iz debla Bacteroidetes. Opredelitev mikrobne združbe bakterijskih zobnih oblog, biofilma iz subgingivalnega prostora pacientov s parodontalno boleznijo kaže na pomen bakterij iz debla Bacteroidetes, kot ključnih v sami patogenezi bolezni. V klinični praksi se kaže potreba po razvoju robustnih diagnostičnih metod v začetnih fazah bolezni. Ena izmed potencialnih molekul, ki bi lahko predstavljala biološki označevalec za odkrivanje parodontalne bolezni je lipid CPE. Zaradi specifične in močne interakcije med egerolizini in CPE, kaže ta vrsta proteinov izjemen potencial kot molekularno orodje v diagnostiki bolezni, ki je sicer povezana z višjimi ravni lipida CPE. S tem namenom smo z uporabo erilizina A z metodama tankoplastne kromatografije in točkovnim nanosom lipidov kvalitativno opredelili prisotnost lipida CPE v kliničnih vzorcih posameznikov s parodontalno boleznijo ter pri parodontalni bakteriji Porphyromonas gingivalis. Hipotezo o prisotnosti CPE pri bakteriji P. gingivalis smo z natančno lipidomsko analizo potrdili s tandemsko masno spektrometrijo z elektrorazpršilno ionizacijo (LC-ESI-MS/MS) in jedrsko magnetno resonanco (NMR).

Language:Slovenian
Keywords:egerolizini, parodontalna bolezen, ceramid fosfoetanolamin, EryA, molekularni označevalci
Work type:Master's thesis/paper
Typology:2.09 - Master's Thesis
Organization:BF - Biotechnical Faculty
Place of publishing:Ljubljana
Publisher:[H. Munjaković]
Year:2021
PID:20.500.12556/RUL-133308 This link opens in a new window
UDC:579.61:616.311:577.112:577.2.088
COBISS.SI-ID:85842947 This link opens in a new window
Publication date in RUL:20.11.2021
Views:879
Downloads:39
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Secondary language

Language:English
Title:Aegerolysin-like proteins as new diagnostic tools for detection of ceramide phosphoetnaolamine, a biomarker of periodontal disease-associated bacteria
Abstract:
Aegerolysins are small (~13-20 kDa), β-structured proteins, with common prominent feature to interact with specific lipids / lipid domains, that are building blocks of various biological membranes. Some fungal aegerolysins are known to strongly associate with ceramide phosphoetanolamine (CPE) containing membranes. Ceramide phosphoetanolamine is characteristic lipid for prokaryotic phylum Bacteroidetes. Research in microbiome of dental plaque in periodontal disease has shown that Bacteroidetes are highly abundant in subgingival plaque samples of chronic periodontitis patients and represent the main causative bacterial agents of periodontal disease. To date, no accurate, precise, and reproducible indicators of periodontal disease have been developed. One of the potential molecules that can be considered as a biomarker of periodontal disease is a specific sphingolipid molecule, ceramide CPE. Due to the specific and strong interaction between aegerolysins and CPE, these proteins have potential to be used as molecular tools for the diagnosis of various diseases associated with higher levels of CPE. In this master thesis specific binding of erylysin A to CPE in lipid extracts of the periodontal bacteria Porphyromonas gingivalis and clinical samples of individuals diagnosed with periodontitis using dot blot and thin layer chromatography analyses was shown. Hypothesis of presence CPE in bacteria P. gingivalis was confirmed with precise lipidomic analysis carried out using liquid chromatography – electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) and nuclear magnetic resonance (NMR).

Keywords:aegerolysins, periodontal disease, ceramid phosphoetanolamin, EryA, biological markers

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