20.500.12556/RUL-149695 Določanje termodinamske stabilnosti riboza-fosfat pirofosfokinaze iz Escherichie coli Thermodynamics stability of Ribose-phosphate pyrophosphokinase from Escherichia coli Encim riboza-fosfat pirofosfokinaze ali PRPP sintetaza katalizira pretvorbo riboze-5-fosfat v fosforibozil pirofosfat, ki je prekurzor za sintezo purinskih in pirimidinskih nukleotidov. Gre za esencialen encim, ki je dobro ohranjen in ga najdemo tako pri bakterijskih organizmih kot pri človeku. V diplomskem delu smo izvedli kemijsko denaturacijo sečnine in le-to spremljali s CD spektroskopijo. CD spektri kažejo obliko značilno za alfa-helični protein, in da se pri visoki koncentraciji sečnine poruši večina njegove sekundarne strukture. Denaturacija s sečnino je reverzibilna. Denaturacijske krivulje določene s CD spektroskopijo ne kažejo odvisnosti od koncentracije proteina. Z analizo denaturacijskih krivulj smo določili termodinamsko stabilnost PRPP sintetaze (standardna Gibbsova prosta energija denaturacije) ki znaša 9,7 kJ/mol. The enzyme ribose-phosphate pyrophosphokinase or PRPP synthetase catalyzes the conversion of ribose-5-phosphate to phosphoribosyl pyrophosphate, which is a precursor for the synthesis of purine and pyrimidine nucleotides. It is an essential enzyme that is well conserved and is found in both bacterial organisms and humans. In the thesis, we carried out the chemical denaturation of urea and monitored it with CD spectroscopy. The CD spectra show characteristic of alpha-helical protein, and that at high concentration of urea most of its secondary structure is destroyed. Denaturation by urea is reversible. Denaturation curves determined by CD spectroscopy show no dependence on protein concentration. By analyzing denaturation curves, we determined the thermodynamic stability of PRPP synthetase (standard Gibbs free energy of denaturation), which amounts to 9,7 kJ/mol. denaturacija PRPP sintetaza termodinamska stabilnost CD spektroskopija denaturation PRPP sythetaze thermodynamic stability CD spectroscopy true false false Slovenski jezik Angleški jezik Diplomsko delo/naloga 2023-09-08 10:45:00 2023-09-08 10:45:04 2023-10-20 13:01:43 0000-00-00 00:00:00 2023 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 22669 169295107 Popovic_Spela_-_Dolocanje_termodinamske_stabilnosti_riboza-fosfat_pirofosfokinaze_iz_Escherichie.pdf Popovic_Spela_-_Dolocanje_termodinamske_stabilnosti_riboza-fosfat_pirofosfokinaze_iz_Escherichie.pdf 1 E38C54D665020B8A92F4CBA3ECB67C46 2397df79eee48d6b9d928c8a4097a334af3c231f144643dd34da530c63c9b7c4 01c7b735-4e24-11ee-9323-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=174190 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0