This thesis investigated the avian reovirus sigma NS protein and its interaction with RNA using mass spectrometric methods. The sigma NS structure is still not known; thus, hydrogen-deuterium exchange and cross-linking coupled to mass spectrometry were employed to understand protein conformation. This research provides insights into protein-RNA interactions and the obtained data will be used in future work to predict protein structure.
Anotace v angličtině
This thesis investigated the avian reovirus sigma NS protein and its interaction with RNA using mass spectrometric methods. The sigma NS structure is still not known; thus, hydrogen-deuterium exchange and cross-linking coupled to mass spectrometry were employed to understand protein conformation. This research provides insights into protein-RNA interactions and the obtained data will be used in future work to predict protein structure.
Klíčová slova
Avian reovirus, sigma NS protein, 3D structure, matrix-assisted laser/desorption ionization, hydrogen-deuterium exchange, cross-linking mass spectrometry.
Klíčová slova v angličtině
Avian reovirus, sigma NS protein, 3D structure, matrix-assisted laser/desorption ionization, hydrogen-deuterium exchange, cross-linking mass spectrometry.
Rozsah průvodní práce
46 p.
Jazyk
AN
Anotace
This thesis investigated the avian reovirus sigma NS protein and its interaction with RNA using mass spectrometric methods. The sigma NS structure is still not known; thus, hydrogen-deuterium exchange and cross-linking coupled to mass spectrometry were employed to understand protein conformation. This research provides insights into protein-RNA interactions and the obtained data will be used in future work to predict protein structure.
Anotace v angličtině
This thesis investigated the avian reovirus sigma NS protein and its interaction with RNA using mass spectrometric methods. The sigma NS structure is still not known; thus, hydrogen-deuterium exchange and cross-linking coupled to mass spectrometry were employed to understand protein conformation. This research provides insights into protein-RNA interactions and the obtained data will be used in future work to predict protein structure.
Klíčová slova
Avian reovirus, sigma NS protein, 3D structure, matrix-assisted laser/desorption ionization, hydrogen-deuterium exchange, cross-linking mass spectrometry.
Klíčová slova v angličtině
Avian reovirus, sigma NS protein, 3D structure, matrix-assisted laser/desorption ionization, hydrogen-deuterium exchange, cross-linking mass spectrometry.
Zásady pro vypracování
-
Zásady pro vypracování
-
Seznam doporučené literatury
-
Seznam doporučené literatury
-
Přílohy volně vložené
-
Přílohy vázané v práci
ilustrace, grafy, tabulky
Převzato z knihovny
Ne
Plný text práce
Přílohy
Posudek(y) oponenta
Hodnocení vedoucího
Záznam průběhu obhajoby
Defence was hybrid, the student was present in person, the members of the commission were present in person or connected via MS Teams.
Prof. Grubhoffer welcomed the student and the commission.
The student presented his work „Mass spectrometry in structural proteomics - study of viral protein complexes “, introduction to the topic, methods, results, summary and future plans.
Supervisor Dr. Dyčka presented his review.
Oponnent Dr. Loginov presented his review.
Student answered opponent´s questions on error of MALDI measurement, peaks mass, cross-linking SDS-PAGE gel, cross-linking optimization, differences between intra- and inter-molecular cross-links, correct structure prediction, the role of SUMO part, buffer composition for quenching reactions in HDX experiments, addition of reduction agents, impact of unreduced Cys on HDX data, digestion conditions, different proteases, two forms of the protein in HDX data, error of HDX measurement, data normalization, apperance of Ex1 kinetics in the deuterium uptake upon RNA binding.
Student answered questions of the members of the commission:
Mgr. Koník – ideas for improvements of MALDI experiments