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Biološka vloga interakcije bakterijskih egerolizinov z lipidi
ID Kočar, Eva (Author), ID Butala, Matej (Mentor) More about this mentor... This link opens in a new window

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Abstract
Protein RahU, ki ga sintetizira bakterija Pseudomonas aeruginosa, uvrščamo v proteinsko družino egerolizinov (Pfam 06355), ki vključuje predstavnike predvsem iz gliv in bakterij. Skupna lastnost glivnih in bakterijskih egerolizinov je njihova vezava na membranske lipide, nekateri izmed njih pa vsebujejo hemolitično in toksično aktivnost proti evkariontskim celicam. RahU veže oksidiran lipoprotein nizke gostote, lizofosfatidilholin ter bakterijske biosurfaktante ramnolipide. Z namenom prepoznave biološke pomembnosti interakcije egerolizina RahU z lipidi, smo uvedli aminokislinske zamenjave v okolici amino-terminalnega konca proteina RahU in s pomočjo heterolognega ekspresijskega sistema Escherichia coli pridobili rekombinanten protein RahU ter njegove različiče. Dokazali smo, da omenjene zamenjave ne vplivajo na pravilno zvitje in stabilnost mutantov. S testom sedimentacije multilamelarnih veziklov in površinsko plazmonsko resonanco smo pokazali vezavo RahU s ceramid fosfoetanolaminom, poglavitnim sfingolipidom v membranah nevretenčarjev. RahU je tako prvi znani bakterijski egerolizin, ki veže omenjeni lipid. Prav tako smo pokazali vezavo RahU z O-fosforiletanolaminom ter nevroendokrinim hormonom noradrenalinom, testi rojenja sevov bakterije Pseudomonas aeruginosa PA01, PW1205 in PW1206 pa so pokazali, da egerolizin RahU oslabi rojenje teh bakterij.

Language:Slovenian
Keywords:egerolizini, Pseudomonas aeruginosa, RahU, ceramid fosfoetanolamin, rojenje
Work type:Master's thesis/paper
Organization:BF - Biotechnical Faculty
Year:2019
PID:20.500.12556/RUL-107612 This link opens in a new window
COBISS.SI-ID:5068623 This link opens in a new window
Publication date in RUL:07.05.2019
Views:1903
Downloads:576
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Secondary language

Language:English
Title:Biological role of bacterial aegerolysin interaction with lipids
Abstract:
Aegerolysin RahU of the Pseudomonas aeruginosa is a bacterial protein belonging to the aegerolysin protein family (Pfam 06355), mainly composed of proteins originating from fungi and bacteria. The common property of fungal and bacterial aegerolysins is their ability to interact with membrane lipids. Some of them also possess hemolytic and toxic activity against eukaryotic cells. The RahU interacts with oxidized low-density lipoprotein, lysophosphatidylcholine and bacterial biosurfactants rhamnolipids. To elucidate the biological importance of the interaction between aegerolysin RahU and lipids, we successfully purified single alanine substitution RahU mutants in the Escherichia coli heterologous expression system. We proved that single amino acid substitutions did not affect the structure and the stability of the RahU derivatives. With sedimentation assay and surface plasmon resonance spectrometry, we showed that RahU interacts with ceramide phosphoethanolamine, the main sphingolipid in the membranes of invertebrates. Therefore, this is the first report of a bacterial aegerolysin that interacts with the aforementioned lipid. We have also shown the binding of RahU to O-phosphorylethanolamine and the neuroendocrinic hormone noradrenaline. In addition, by using the swarming tests on Pseudomonas aeruginosa PA01, PW1205 in PW1206 strains, we suggest that aegerolysin RahU affects the swarming of Pseudomonas aeruginosa.

Keywords:aegerolysins, Pseudomonas aeruginosa, RahU, ceramide phosphoethanolamine, swarming

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