Autor: Urban, M.; Soural, M.; Hodon, J.; Sidova, V.; Sarek, J.; Borkova, L.; Piskorova, K.; Rehulka, J.; Dzubak, P.; Hajduch, M.
Název práce v češtině: Biotinylované triterpeny a jejich využití při hledání cílů
Název práce v angličtině: Biotinylated Triterpenes and Their Use in Target Identification
Klíčová slova v češtině: triterpeny, biotin, mechanizmus účinku, cíle
Klíčová slova v angličtině:triterpenes, biotin, mechanism of action, targets
Abstrakt česky: Pro lepší pochopení mechanizmu účinku cytotoxických triterpenů jsme se rozhodli zkoumat jejich molekulární cíle. Připravili jsme set biotinylovaných sloučenin a pomocí pull down esejí s využitím kvantitativní proteomiky a SILAC identifikujeme proteiny, se kterými vytváří nekovalentní interakci
Abstrakt anglicky: To better understand the mechanism of action of antitumor triterpenes, we are trying to identify their molecular targets. Pull down assays that use combination of quantitative proteomics with SILAC seem to be the most efficient since they can identify many targets in one experiment. The method uses compounds anchored to magnetic beads via biotin-straptavidin interaction. For that, the active derivative needs to be connected to biotin through a defined linker. In this work, we developed a simple and fast solid-phase synthetic approach to connect terpenes with biotin. Terpenes were biotinylated from three different sides (C-3, C-28, C-30, see formula 1) which should help to better characterize their pharmacophore. We expect to isolate the target protein(s) in case, when the molecule is connected to biotin through its side away from the pharmacophore while we expect negative result when biotinylation was made through the pharmacophore and hinders it. A set of seven triterpenoid derivatives of high cytotoxicity were biotinylated (for example of a compound biotinylated through C-3 see formula 2). The cytotoxicity was similar to the non-biotinylated parents and this indicates that the target identification should not be thwarted by the presence of the linker and biotin. Our research is now focused on pull down assays optimization, target validation and evaluation of various linkers. The developed solid-phase synthetic approach1 is the first attempt to use solid phase synthesis to connect active triterpenes to biotin and is applicable as a general procedure for routine preparation of conjugates of triterpenes with various molecules. Synthesis, pull down experiments and examples of target proteins will be shown.
Jazyk v originále: Anglický
Publikace abstraktu: Sborník 151/166
Forma prezentace: poster
Název konference: Advances in Organic, Bioorganic and Pharmaceutical Chemistry „Liblice 2015“
Místo konání: Olomouc (Czech Republic)
Datum konání: November 6-8
Rok konání: 2015
Způsob financování: GACR 15-05620S