Poništavanje hepatoprotektivnog učinka D-met-enkefalina komplementarnim peptidom kao model za proučavanje peptidnih interakcija

master's thesis

• Title (english): Reversal of the hepatoprotective effect of D-met-enkephalin by a complementary peptide as a model for studying peptide interactions
• Author: Katarina Matković
• Mentor: Paško Konjevoda (mentor)
• Mentor: Rosa Karlić (sumentor)
• Committee member: Rosa Karlić (predsjednik povjerenstva)
• Committee member: Duje Lisičić (član povjerenstva)
• Committee member: Martina Šeruga Musić (član povjerenstva)
• Committee member: Sofia Ana Blažević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2021-02-26, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Većina regulatornih procesa u biološkim sustavima ovisi o interakcijama između proteina. Naše
spoznaje o takvim interakcijama su relativno ograničene, a eksperimentalne metode za njihovo
proučavanje kompleksne, spore i skupe. Zbog toga se u radu opisuju heurističke metode za
određivanje bioaktivnih područja u proteinima te interakcije tih područja s drugim proteinima.
Jedna od takvih metoda, teorija molekularnog prepoznavanja (MRT) praktično je primijenjena u
biološkom eksperimentalnom modelu poništenja učinka peptida D-met-enkefalina. Protektivni
učinak D-met-enkefalina proučavan je u in vivo modelu oštećenja jetre mužjaka miša CBA soja
paracetamolom uz primjenu tri kriterija: aktivnost enzima ALT i AST, patohistološko određivanje
stupnja nekroze jetre te broj spontano uginulih životinja. Dokazan je protektivni učinak po dva
kriterija: aktivnost enzima ALT (U/L) i AST (U/L) te bodovi nekroze jetre. Optimalna protektivna
doza iznosi 5 mg/kg. Protektivni učinak poništavao se uz pomoć komplementarnog peptida (IPPLY)
konstruiranog uporabom MRT. Osnovna svojstva komplementarnog peptida te njegova interakcija s
D-met enkefalinom proučavana su pomoću cirkularne dikroizmatske spektroskopije i fluorescentne
spektroskopije. Antisens peptid u potpunosti poništava protektivni učinak optimalne doze D-metenkefalina.
Dokazano je da je uporabom teorije molekularnoga prepoznavanja moguće na
jednostavan i učinkovit način izraditi peptidni sustav liganda i akceptora.

Abstract (english)

Most regulatory processes in biological systems depend on protein-protein interactions. Our
knowledge of such interactions is relatively limited, and experimental methods for studying them
are complex, slow and expensive. Therefore, heuristic methods to determine bioactive regions in
proteins and interactions of these regions with other proteins are described in the paper. One such
method, Molecular Recognition Theory (MRT), has been practically applied in the biological
experimental model of reversal effect of the D-met-enkephalin peptide. Protective potential of Metenkephalin
was evaluated using an experimental model of paracetamol induced hepatotoxicity in
male CBA mice and observed by using three criteria: plasma activities of ALT and AST enzymes,
liver necrosis score and number of spontaneously dead animals. Protective potential was
demonstrated by two criteria: activity of enzymes ALT (U / L) and AST (U / L) and liver necrosis
score. The optimal protective dose is 5 mg / kg. Protective effect was reversed using a
complementary peptide (IPPLY) constructed using MRT. The basic properties of the
complementary peptide and its interaction with D-met enkephalin were studied using circular
dichroism spectroscopy and fluorescence spectroscopy. The antisense peptide completely reverses
the protective effect of the optimal dose of D-met-enkephalin. It has been proven that using
molecular recognition theory it is possible to construct a ligand-acceptor peptide system in a simple
and efficient way.

• Keywords: heuristički algoritmi; teorija molekularnoga prepoznavanja; komplementarni peptidi. metenkefalin; hepatoprotekcija
• Keywords (english): heuristic algorithms; molecular recognition theory; complementary peptides; met-enkephalin; hepatoprotection
• Language: croatian
• URN:NBN: urn:nbn:hr:217:811245
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access; Embargo expiration date: 2023-02-26
• Created on: 2021-03-16 12:17:17