Role dvou lysinů v transmembránové doméně lidského P2Y12 receptoru a vliv na vazbu agonistů

• Main Author: Ćwierz, Jan
• Other Authors: Pávek, Petr
• Format: Dissertation
• Language: English
• Published: 2011
• Online Access: http://www.nusl.cz/ntk/nusl-312586

Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmacology and Toxicology Candidate: Jan Ćwierz Supervisors: Prof. Dr. med. Ivar von Kügelgen Doc. PharmDr. Petr Pávek, Ph.D. Title of diploma thesis: Role of two lysine amino acid residues in transmembrane regions in agonist recognition of the human P2Y12 receptor Human P2Y12 receptor expressed on platelets plays a key role in their physiological function. It importantly participates in maintenance of normal haemostasis and is often targeted by antiplatelet medication, such as clopidogrel. Replacement of amino acid residues responsible for ADP binding, which is a naturally occurring negatively charged agonist, can lead to impaired ligand-receptor interaction. Aim of this work was to study the role of two lysine amino acid residues in the position 173 and 174 of the P2Y12 receptor protein and subsequent changes in 2-methylthio-ADP binding. Two positively charged lysine amino acid residues were substituted with neutral residue alanine. Receptor expression in Chinese hamster ovary was analyzed by immunofluorescence staining and laser scanning confocal microscopy. Subsequently, the wild-type receptor and three mutant variants (K173A, K174A and K173A/K174A) were examined by means of 2-methylthio-ADP-induced responses in...