Browsing by Subject "fosforylacja"

Now showing 1 - 2 of 2

Access status: Restricted ,
Porównanie różnych technik fragmentacji w spektrometrii mas do analizy fosfopeptydów
(Data obrony: 2021-01-28) Kuchnia, Karolina
Wydział Elektrotechniki, Automatyki, Informatyki i Inżynierii Biomedycznej
Access status: Restricted ,
Quantitative analysis of phosphoproteome of the selected brain structures in morphine dependence
(Data obrony: 2013) Sucharski, Filip
Wydział Inżynierii Materiałowej i Ceramiki
The aim of the project was the search for, and identification of the markers of drug dependence. The research was focused on the identification of molecular mechanisms involved in these processes and clarification of the role of particular proteins in morphine dependence. Project puts special emphasis on phosphoproteins, and in general on phosphorylation, as a main mechanism involved in control of essentially any biological process and many diseases. High dynamism and reversible character, make phosphorylation a main regulator of signaling networks. Therefore, a global wide-scale analysis of phosphoproteome however, troublesome, gives hope on detailed recognition of complicated molecular mechanisms and mutual connections between metabolic pathways. It is known, that drug dependence has a huge impact on many cell functions as well as biological processes, for which the relation with the drug dependence seems to be indirect. Thus, only a global analysis of a state of phosphoproteins, in particular kinases with their mutual relations, can contribute to solving a puzzle. The project involves the newest, fully quantitative, methods for phosphoproteome analysis. In particular a special work-flow based on two methods of phosphopeptides enrichment (TiO2, IMAC), data- dependent mass spectrometry acquisition and specific bioinformatic platform were developed. This approach allowed for simultaneous analysis of CID/ETD data from the Bruker ion-trap mass spectrometer with Mascot as identification tool and finally with the Trans-proteomic pipeline. This work describes methods for phosphoproteomics and gives a detailed protocols for sample preparation, phosphopeptides enrichment and mass spectrometry analysis. However it puts special emphasis on data analysis including self-programmed python scripts for data conversion, which is crucial in such wide-scale experiments. The project resulted in identification and quantitation of a large number of phosphoproteins, which may be considered as potential biomarkers for morphine dependence. A few of them were discussed in details, however further research, especially based on MRM (Multiple reaction monitoring) approach is required.