Utjecaj promjene glikozilacije humanog serumskog transferina na otpuštanje željeza u prisutnosti kompetitivnog kelatora EDTA

master's thesis

• Title (english): Effect of change in glycosylation of human serum transferrin on iron release to competing chelator EDTA
• Author: Helena Deriš
• 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: 2017-11-23, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

U ovom radu proučavan je utjecaj promjene glikozilacije transferina na konstantu vezanja željeza. Svrha rada bila je dokazati utjecaj glikanskog dijela strukture transferina na njegovu funkciju, točnije, na jačinu vezanja željeza. Sva ispitivanja su se provodila u PIPES puferu pri pH 7.4, na temperaturi 25°C, a kompleksirajući kompetitivni agens bila je EDTA. Korištena tehnika bila je spektroskopija u UV-Vis području. Iz podataka o apsorbanciji na 450 nm i koncentraciji dodane EDTA, izračunate su najprije pripadajuće koncentracije apo-transferina i holo-transferina te koncentracija otpuštenog željeza, a zatim i odgovarajuće konstante ravnoteže. Dobivene konstante ravnoteže veće su za red veličine kod vezanja željeza za nativni transferin u odnosu na asijalo-transferin. Također, kinetika otpuštanja željeza s asijalo-transferina je promijenjena u odnosu na nativnu sijaliniziranu formu.

Abstract (english)

This work investigates the effect of change of transferrin glycosylation on the iron binding constant. The purpose of this study was to demonstrate the influence of the glycan part of the transferrin structure on its function, specifically on the iron binding strength. All experiments were carried out by UV-Vis spectroscopy in PIPES buffer (pH = 7.4) at 25° C and EDTA was used as a competitive chelator. Based on the absorbance data at 450 nm and the concentration of the added EDTA, concentrations of apo-transferin, holo-transferin and free iron were first obtained and then the corresponding equilibrium constants were calculated. The obtained equilibrium constants are an order of magnitude higher for the iron binding to the native transferrin, compared to asialo-transferrin. Additionally, the iron release kinetics from asialo-transferrin has been altered relative to the native sialylated protein form.

• Keywords: transferin; željezo; glikozilacija; desijalinizacija; EDTA
• Keywords (english): transferrin; iron; glycosylation; desialylation; EDTA
• Language: croatian
• URN:NBN: urn:nbn:hr:163:525460
• 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
• Created on: 2018-01-24 16:20:29