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Titre : | Study of the potential oligomerization of peroxiredoxins in living cells by monitoring their activity over time in Saccharomyces cerevisiae |
Auteurs : | Antoine Demoulin, Auteur ; B. MORGAN, Promoteur |
Type de document : | Travail de fin d'études |
Editeur : | Woluwe-Saint-Lambert : Haute École Léonard de Vinci, 2021 |
Langues: | Anglais |
Index. décimale : | TFE - Biologie médicale |
Descripteurs : |
HE Vinci Biochimie ; Biotechnologie |
Mots-clés: | eucaryotes |
Résumé : |
Peroxiredoxins (Prxs) are enzymes reducing chemical compounds named peroxide. High peroxide concentrations are considered as a threat to living cells due to their toxicity. It can cause several diseases such as diabetes, Alzheirmers disease or cancer. It is known that Prxs form dimers and then decamers. Eukaryotes organisms produce several similar Prxs in different cellular compartments.
Even though PubMed published more than 5000 publications about Prxs, only two did consider the possible formation of heterodecamers. Every previous publication considered that decamers were formed with ten same Prxs. Therefore, we decided to use genetically encoded fluorescent probes to help us to discover if other Prxs form heterodecamers. The probe is composed of the Prx on which we want to obtain information, linked to a fluorescent molecule. By inserting genetic patrimony inside Saccharomyces cerevisiae cells, we have the fluorescent probe produced by the yeast. Furthermore, once the yeast cells are transformed, we monitor the in vivo activity of these Prxs over time to observe if a heterodimer is formed between the probe and an endogenous Prx. This will be done by adding hydrogen peroxide to the wells containing the yeast cells, in a fluorometer. Fluorescence will then be measured and depending on the reactivity of the Prx, the intensity of the excitation wavelength spectrum will be influenced. It came out that several Prxs analyzed do form heterodimers. This could have an influence on the functions fulfilled by these enzymes like signaling a different information or having diverse chaperone functions. Besides that, this experiment informed us about the sensitivity of the probes used. When the RoGFP2-Tsa2 ΔCR probe was created in 2016, it was considered as the most sensitive sensor to monitor basal H2O2 levels in yeast cells. We compared it to other probes used and we observed that roGFP2-AtPrdxA ΔCR is even more sensitive. This gives good hope for the future because it could respond to lower H2O2 ranges. |
Accès : | Identifiez-vous avant d'accéder au document électronique |
Disponible en ligne : | Oui |
Lieu du stage : | Universität des Saarlandes, Saarbrücken, Germany |
Département : | Biologie médicale |
Documents numériques (1)
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TFE Biologie médicale Adobe Acrobat PDF |