20.500.12556/RUL-182297 Uporaba 4-okso-4H-piridino[1,2-a]pirimidin-3-diazonijevega in 1-ciano-4-okso-4H-kinolizin-3-diazonijevega tetrafluoroborata v fluorescenčnem označevanju proteinov Application of 4-oxo-4H-pyridino[1,2-a]pyrimidine-3-diazonium and 1-cyano-4-oxo-4H-quinolizine-3-diazonium tetrafluoroborate in fluorescent labeling of proteins 4-okso-4H-piridino[1,2-a]pirimidin–3–diazonijev in 1-ciano-4-okso-4H-kinolizin–3– diazonijev tetrafluoroborat sta bila izbrana kot reagenta za izvedbo fotokemijskih sulfanilacij in arilacij ter termičnih sulfanilacij z N-Boc-zaščitenimi estri cisteina in cistina ter aromatskimi aminokislinami. Raziskali smo stabilnost obeh izhodnih reagentov v vodnem mediju z namenom nadaljnje vezave (aza)kinolizinskih diazonijevih soli na proteine. Obe diazonijevi soli sta bili uspešno uporabljeni v reakcijah biokonjugacije. Dokazali smo, da je (aza)kinolizin diazonijeve tetrafluoroborate mogoče uporabiti pri fluorescentnem označevanju proteinov, saj se pod fotokataliziranimi pogoji vežejo tako na tiolne skupine in disulfidne vezi cisteinskih ostankov, kot tudi na arilne skupine aromatskih aminokislin. Pod termičnimi pogoji pa je prednostna reakcija sulfaniliranja (vezava na tiolne skupine cisteinskih preostankov). Kot modelni protein je bil izbran goveji serumski albumin (BSA). Razvili smo tudi metodo za kvantifikacijo vezave in optimizirali pogoje vezave. 4-Oxo-4H-pyridino[1,2-a]pyrimidine–3–diazonium and 1-cyano-4-oxo-4H-quinolizine–3–diazonium tetrafluoroborate were selected as starting reagents for photochemical sulfanylations and arylations, as well as thermal sulfanylations with esters of N-Boc-protected cysteine, cystine and aromatic amino acid. The stability of both starting reagents in aqueous media was investigated to enable further coupling of (aza)quinolizine diazonium salts to proteins. Both diazonium salts were successfully used in bioconjugation reactions. We demonstrated that (aza)quinolizine diazonium tetrafluoroborates can be used for fluorescent labeling. Under photocatalyzed conditions, these diazonium salts bind to thiol and disulfide bonds of cysteine residues, as well as to aryl groups of aromatic amino acid residues. In contrast, the sulfanylation reaction is favored under thermal conditions. Bovine serum albumin (BSA) was selected as a model protein. We also developed a method for quantifying binding and optimized binding conditions. diazonijeve soli sulfanilacije arilacije fotoredoks kataliza biokonjugacija fluorescentno označevanje diazonium salts sulfanylations arylations photoredox catalysis bioconjugation fluorescent labeling true false false Slovenski jezik Angleški jezik Magistrsko delo/naloga 2026-05-06 14:40:03 2026-05-06 14:40:18 2026-05-07 04:53:20 0000-00-00 00:00:00 2026 0 0 0000-00-00 NiDoloceno NiDoloceno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 1970-01-01 25715 Groselj_Lena_-_Uporaba_4-okso-4H-piridino[1,2-a]pirimidin-3-diazonijevega_in_1-ciano-4-okso-4H-k.pdf Groselj_Lena_-_Uporaba_4-okso-4H-piridino[1,2-a]pirimidin-3-diazonijevega_in_1-ciano-4-okso-4H-k.pdf 1 B2F52FD29BFDDB537B0AA7E279A8EC1B e4c9f9e408c9406f9c785cbbc4eb89f0691830b932e3ea525d27663d67e6733b 742652a5-4948-11f1-b0ab-0050569b8976 https://repozitorij.uni-lj.si/Dokument.php?lang=slv&id=233436 Fakulteta za kemijo in kemijsko tehnologijo 0 0 0