Testing the interaction of flavonoids with protein disulfide isomerase PDIA3 and the effects on protein biological properties

master's thesis

• Author: Ines Kanski
• Mentor: Olga Gornik (mentor)
• Committee member: Olga Gornik
• Committee member: Sandra Šupraha Goreta
• Committee member: Suzana Inić
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of biochemistry and molecular biology); Zagreb
• Defense date and country: 2015-09-14, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

U okviru ovog diplomskog rada istražena je interakcija između odabranih flavonoida (eupatorin, apigenin, apigenin-7-glukozid, kempferol, naringenin, 3-O-metilkvercetin, cijanidin, genistein, luteolin-7-glukozid) sa proteinom ERp57 koji spada u skupinu disulfid-izomeraza (PDI). Za procjenu vezanja flavonoida sa proteinom ERp57 korištena je fluorescencijska spektrofotometrija i učinak gašenja fluorescencije, dok je za ispitivanje utjecaja flavonoida na vezanje ERp57-DNA korištena migracijska elektroforeza (EMSA). Također je testiran utjecaj flavonoida na aktivnost i stabilnost proteina. Apigenin i eupatorin pokazali su najmanju konstantu disocijacije, odnosno najveći afinitet za ERp57 te ih to čini najboljim molekulama za vezanje na protein ERp57. Potvrđena je važnost hidroskilne skupine za interakciju flavonoida sa proteinom.

Abstract (english)

In this thesis, interaction between a protein that belongs to the family of protein disulfide isomerases (PDI)- protein ERp57, and chosen flavonoids- eupatorin, apigenin, apigenin-7-glucoside, kaempferol, naringenin, 3-O-Methylquercetin, cyanidin, genistein and luteolin-7-glucoside was tested. Interaction between ERp57 and flavonoids was assessed using fluorescence quenching method, while electrophorectic mobility shift assay (EMSA) was used to evaluate flavonoid effect on ERp57-DNA interaction. Flavonoid influence on protein activity and protein stability was also tested. Apigenin and eupatorin showed the lowest dissociation constant and the highest binding affinity to ERp57, making them the best compounds to be used as selective inhibitors/modulators of ERp57 biological activities.

• Keywords: flavonoidi; apigenin; eupatorin; ERp57; gašenje fluorescencije
• Keywords (english): flavonoids; apigenin; eupatorin; ERp57; fluorescence quenching
• Language: english
• URN:NBN: urn:nbn:hr:163:378527
• Study programme: Title: Pharmacy; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Repository: Repository of Faculty of Pharmacy and Biochemistry University of Zagreb
• Created on: 2016-01-22 15:11:55