Phosphoproteomic study of molecular Phosphoproteomic study of molecular

mechanismsmechanisms of drug dependenceof drug dependenceF. SucharskiF. Sucharski11, P.Suder, P.Suder11, G.Schroeder, G.Schroeder22, J.Silberring, J.Silberring1,31,3

11 Department of Biochemistry and Neurobiology, Faculty of Materials Sciences and Ceramics, AGH University of Science and Technology, Mickiewicza 30 Ave, 30-059 Krakow, Poland Department of Biochemistry and Neurobiology, Faculty of Materials Sciences and Ceramics, AGH University of Science and Technology, Mickiewicza 30 Ave, 30-059 Krakow, Poland22 Department of Chemistry, Adam Mickiewicz University, Poznan, Poland Department of Chemistry, Adam Mickiewicz University, Poznan, Poland33 Center of Polymer and Carbon Materials, Polish Academy of Sciences, Sowinskiego 5 Str, 44-121 Gliwice, Poland Center of Polymer and Carbon Materials, Polish Academy of Sciences, Sowinskiego 5 Str, 44-121 Gliwice, Poland

IntroductionIntroductionHigh dynamism and reversible character, make phosphorylation a main regulator of signaling networks. Therefore, a global wide-scale analysis of 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 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 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 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.with their mutual relations, can contribute to solving a puzzle.The project is pioneer work in neuroscience, involving the newest, fully quantitative, methods for phosphoproteome analysis. In particular, The project is pioneer work in neuroscience, involving the newest, fully quantitative, methods for phosphoproteome analysis. In particular, phosphoproteome analysis is based on the methods for phosphopeptides isolation, and mass spectrometry as an analytical tool for identification phosphoproteome analysis is based on the methods for phosphopeptides isolation, and mass spectrometry as an analytical tool for identification and quantification. This approach consists in identification of all possible phosphoproteins, specific peptides fragmentation method, and annotation and quantification. This approach consists in identification of all possible phosphoproteins, specific peptides fragmentation method, and annotation of phosphorylation sites.of phosphorylation sites. Here we present a workflow for quantitative phosphopeptides analysis and preliminary results from phosphoproteomic study of prefrontal cortices of morphine dependent rats.