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master's thesis
Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom
Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry, 2022. urn:nbn:hr:163:404231
Atelj, Tamara
University of Zagreb
Faculty of Pharmacy and Biochemistry
Department of general and inorganic chemistry

Institutional repository: Repository of Faculty of Pharmacy and Biochemistry University of Zagreb

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Atelj, T. (2022). Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom (Master's thesis). Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry. Retrieved from https://urn.nsk.hr/urn:nbn:hr:163:404231

Atelj, Tamara. "Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom." Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, 2022. https://urn.nsk.hr/urn:nbn:hr:163:404231

Atelj, Tamara. "Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom." Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, 2022. https://urn.nsk.hr/urn:nbn:hr:163:404231

Atelj, T. (2022). 'Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom', Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, accessed 25 November 2024, https://urn.nsk.hr/urn:nbn:hr:163:404231

Atelj T. Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom [Master's thesis]. Zagreb: University of Zagreb, Faculty of Pharmacy and Biochemistry; 2022 [cited 2024 November 25] Available at: https://urn.nsk.hr/urn:nbn:hr:163:404231

T. Atelj, "Ispitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom", Master's thesis, University of Zagreb, Faculty of Pharmacy and Biochemistry, Zagreb, 2022. Available at: https://urn.nsk.hr/urn:nbn:hr:163:404231

Metadata
TitleIspitivanje razlike u hidrofobnosti nativnog i desijaliziranog transferina reakcijom sa 8-anilinonaftalensulfonskom kiselinom
Title (english)Analysis of the difference in hydrophobicity of native and desialylated transferrin by reaction with 8-anilinonapthalenesulfonic acid
AuthorTamara Atelj
MentorTin Weitner (mentor)
Committee memberTin Weitner (predsjednik povjerenstva)
Committee memberMario Gabričević (član povjerenstva)
Committee memberDavor Šakić (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Pharmacy and Biochemistry
(Department of general and inorganic chemistry)
Zagreb
Defense date and country2022-09-09, Croatia
Scientific / art field,
discipline and subdiscipline		BIOMEDICINE AND HEALTHCARE
Pharmacy
Pharmacy
Abstract
U sklopu ovoga diplomskog rada proučavan je utjecaj vezanja hidrofobne probe, 8-anilino-1-naftalensulfonske kiseline, na fluorescenciju humanog nativnog apo-transferina i desijaliniziranog transferina, kao i stehiometrija vezanja kiseline na nativni protein. Ispitivanje hidrofobnosti provedeno je u fosfatnom puferu koncentracije 50 mM, dok je analiza stehiometrije vezanja hidrofobne probe na ljudski apo-transferin provedena u fosfatnom puferu koncentracije 200 mM, a ispitivanja provedena su pri neutralnoj vrijednosti pH i temperaturi od 25°C. Tehnike koje su korištene u eksperimentalnom dijelu rada su UV-Vis spektrofotometrija i fluorescencijska spektroskopija. Rezultati su obrađeni u programu Microsoft Excel. Dodatak ANS otopini ljudskog nativnog apo-transferina uzrokovao je primjetan porast intenziteta fluorescencije, što ukazuje na to da se proba veže za izloženi džep na površini polipeptidnog lanca. Analizom otopine ANS i desijaliziranog ljudskog apo-transferina nije primjećen porast intenziteta fluorescencije, što znači da nije došlo do hidrofobnih interakcija kao u slučaju nativnog apo-transferina. Ispitivanje stehiometrije vezanja ANS i ljudskog nativnog apo-transferina korištenjem Jobove metode ukazuje na to da se proba i protein vežu u omjeru 1:1 i 2:1.
Abstract (english)
As part of this thesis, the influence of binding of the hydrophobic probe, 8-anilino-1-naphthalenesulfonic acid, on the fluorescence of human native apo-transferrin and desialylated transferrin, as well as the stoichiometry of ANS binding to the native protein, were studied. The hydrophobicity test was performed in a phosphate buffer with a concentration of 50 mM, while the analysis of the stoichiometry of binding of the hydrophobic probe to human apo-transferrin was performed in a phosphate buffer with a concentration of 200 mM, and both tests were performed at a neutral value pH and temperature of 25°C. The techniques used in the experimental part of the thesis are UV-Vis spectrophotometry and fluorescence spectroscopy. The results were processed in Microsoft Excel. Addition of ANS to a solution of human native apo-transferrin caused a marked increase in fluorescence intensity, indicating that the probe binds to an exposed pocket on the surface of the polypeptide chain. Analysis of the solution containing the probe and desialylated human apo-transferrin did not show an increase in fluorescence intensity, which indicates that no hydrophobic interactions occurred as was the case with native apo-transferrin solution. Analysis of the stoichiometry of binding of ANS and human native apo-transferrin using Job's plot indicates that the hydrophobic probe and protein bind in a ratio of 1:1 and 2:1.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:163:404231
Study programmeTitle: Pharmacy
Study programme type: university
Study level: integrated undergraduate and graduate
Academic / professional title: magistar/magistra farmacije (magistar/magistra farmacije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2022-09-09 13:04:01