Utjecaj glikozilacije, sinergističkog aniona i pH na vezanje željeza na humani serumski transferin

master's thesis

• Title (english): Influence of glycosylation, synergistic ion and pH on iron binding to human serum transferrin
• Author: Ria Saraga
• Mentor: Tin Weitner (mentor)
• Committee member: Tin Weitner (predsjednik povjerenstva)
• Committee member: Mario Gabričević (član povjerenstva)
• Committee member: Davor Šakić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of general and inorganic chemistry); Zagreb
• Defense date and country: 2020-09-30, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

U sklopu ovog diplomskog rada proučavan je utjecaj promjene glikozilacije transferina, sinergističkog aniona te pH pufera na konstantu vezanja željeza. Ispitivanja su se provodila u MES puferu pri pH 5,6 i temperaturi 25°C uz dodatak karbonata ili oksalata kao sinergijskih aniona. Korištene su tehnike UV-Vis spektrofotometrije i fluorescencije. Za izračun konstanti K1 i K2, koje opisuju nastanak monoferičnog i diferičnog transferina, korišten je program ReactLab™ EQUILIBRIA. Dobivene konstante i titracijske krivulje ukazuju na to da se prvo željezo veže s najvećim afinitetom na sijalizirani transferin u prisutnosti karbonata. Također, željezo se veže većim afinitetom na sijalizirani transferin s karbonatom kao sinergističkim anionom u odnosu na sijalizirani transferin s oksalatom. U prisutnosti oksalata kao sinergističkog aniona željezo se jače veže na desijalizirani transferin. Usporedbom konstanti ravnoteže pri pH 5,6 i pH 7,4 zaključujemo da se željezo jače veže na transferin pri fiziološkom pH. Promjenom glikozilacije, sinergističkog iona te pH, mijena se afinitet vezanja željeza na transferin, a time i koncentracija slobodnog i potencijalno toksičnog željeza u krvi.

Abstract (english)

As part of this thesis, it was investigated how does the change in glycosylation pattern, synergistic anion and pH of a buffer effect iron binding constant. All experiments were carried out by UV-Vis and fluorescence spectroscopy in MES buffer, with carbonate or oxalate, at pH 7.4 and 25° C. The ReactLab ™ EQUILIBRIA program was used to calculate the constants K1 and K2, which describe the formation of monoferric and diferric transferrin. The obtained constants and titration curves indicate that the first iron binds with the highest affinity to sialylated transferrin with carbonate. Also, iron binds with higher affinity to sialylated transferrin with carbonate as a synergistic anion compared to sialylated transferrin with oxalate, and in the presence of oxalate as a synergistic anion, iron binds more strongly to desialized transferrin. By comparing the equilibrium constants at pH 5.6 and pH 7.4, we conclude that iron is more strongly bound to transferrin at physiological pH. By changing glycosylation, synergistic ion and pH, the binding affinity of iron to transferrin changes and thus the concentration of free, potentially toxic, iron in the blood.

• Keywords: transferin; željezo; fluorescencija; UV-Vis spektrofotometrija; konstanta vezanja
• Keywords (english): transferrin; iron; fluorescence; UV-Vis spectroscopy; binding constant
• Language: croatian
• URN:NBN: urn:nbn:hr:163:772009
• 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; Embargo expiration date: 2022-09-30
• Created on: 2022-09-02 12:32:28