magistrsko delo
Eva Kočar (Author), Kristina Sepčić (Reviewer), Matej Butala (Mentor), Matej Butala (Thesis defence commission member), Kristina Sepčić (Thesis defence commission member), Jerneja Ambrožič (Thesis defence commission member)

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.

Keywords

egerolizini;RahU;ceramid fosfoetanolamin;rojenje;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [E. Kočar]
UDC: 577(043.2)
COBISS: 5068623 Link will open in a new window
Views: 1243
Downloads: 532
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Other data

Secondary language: English
Secondary title: Biological role of bacterial aegerolysin interaction with lipids
Secondary 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.
Secondary keywords: aegerolysins;RahU;ceramide phosphoethanolamine;swarming;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. Ljubljana, Biotehniška fak.
Pages: XIII f., 85 str., [9] f.
ID: 11132589