Razvoj metode za mjerenje aktivnosti ekto-ATPaze u serumu čovjeka

master's thesis

• Title (english): Development of method for measuring ecto-ATPase activity in human blood serum
• Author: Monika Janković
• Mentor: Tihana Žanić Grubišić (mentor)
• Committee member: Tihana Žanić Grubišić
• Committee member: Anita Somborac Bačura
• Committee member: Ivan Pepić
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2015-06-08, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Pharmacy

Abstract

Ekto-ATPaza je enzim koji pripada skupini E-NTPDaza. Navedeni enzimi su ektoenzimi, što znači da
sudjeluju u metabolizmu izvanstaničnih molekula. Ekto-ATPaza sudjeluje u metabolizmu nukleozid
difosfata i nukleozid trifosfata, s većim afinitetom prema nukleozid trifosfatima. Metabolizam
izvanstaničnih molekula s odgovarajućim enzimima jedan je od koraka u purinergičnoj signalizaciji koja
pak posreduje u različitim fiziološkim i patofiziološkim procesima. Stoga se smatra da vrijednost aktivnosti
ekto-ATPaze u serumu može biti potencijalni čimbenik u dijagnostici bolesti povezanih s purinergičnom
signalizacijom. Aktivnost ekto-ATPaze u serumu mjerena je pomoću Fiske-Subbarow metode, a srednja
vrijednost izmerena na 13 kontrolnih uzoraka iznosi 992.3±281.2 nmol/ml/h. Rezultati su pokazali da je za
mjerenje aktivnosti najprikladniji volumen uzorka 50 μL, vrijeme inkubacije na 37 °C 60 min te da je
potrebno 100 μL 1.12 mol/L TCA za deproteinizaciju uzorka.

Abstract (english)

Ecto-ATPase is an enzyme which belongs to the group of the E-NTPDases. Those enzymes are ectoenzymes, and that they participate in the metabolism of extracellular molecules. Ecto-ATPase participate in metabolism of nucleoside triphosphate and nucleoside diphosphate with higher affinity for the nucleoside triphosphate. The metabolism of extracellular molecules with the appropriate enzymes is one of the steps in purinergic signaling that mediates a variety of physiological and pathophysiological processes. Therefore it is considered that value of ecto-ATPase activity in human serum may be a potential factor in the diagnosis of diseases associated with purinergic signaling. Ecto-ATPase activity in the human serum was measured by Fiske-Subbarow method, and the average value of 13 healthy samples was 992.3±281.2 nmol/ml/h. Results showed that the most suitable sample volume for the measuring enzyme activity is 50 μL, incubation time on 37 °C is 60 min and that 100 μL 1.12 mol/L TCA is required for deproteinization of serum.

• Keywords: ektoenzimi; ekto-ATPaza; purinergična signalizacija; enzimska aktivnost; Fiske-Subbarow metoda
• Keywords (english): ectoenzymes; ecto-ATPase; purinergic signaling; enzymatic activity; Fiske-Subbarow method
• Language: croatian
• URN:NBN: urn:nbn:hr:163:643723
• 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: 2016-01-18 15:46:50