diplomsko delo
Rebeka Dajčman (Author), Iztok Turel (Mentor)

Abstract

Encimi so pomembne tarče zdravil v boju proti patogenim organizmom in tudi organizmu lastnim motnjam, ki privedejo do bolezenskih stanj. Na prvi pogled se zdi, da proteaze ne sodijo v to skupino encimov, saj le razgrajujejo proteine, kar pa je povsem napačno. Cisteinski katepsini, ki se preferenčno nahajajo v lizosomu, sodelujejo pri veliko več celičnih procesih kot le pri normalni razgradnji zunajceličnega materiala in antigenskem procesiranju. Eden takšnih encimov je katepsin L, ki med drugim sodeluje v razvoju hudega akutnega respiratornega sindroma (SARS). Tudi cisteinski katepsini so postali zanimiva tarča za preučevanje inhibitornega učinka raznih molekul. Eden najbolj raziskanih cinkovih kompleksov je cinkov pirition, ki je že dolgo znan po svojem antimikrobnem delovanju in je ključna komponenta v komercialnih šamponih proti prhljaju. Poleg antimikotičnega delovanja izkazuje tudi protivirusno delovanje, zaradi česar je potencialno zdravilo v boju proti virusnim infekcijam. Le malo je znanega o njegovem učinku na katepsin L, po drugi strani pa so cinkovi ioni znani po tem, da inhibirajo katepsin B, ki prav tako sodi v CA1 družino papainu podobnih proteaz. Iz tega razloga smo se odločili, da preverimo vpliv piritiona v kombinaciji s cinkovimi solmi na aktivnost katepsina L. Najprej smo določili stabilnost vseh testiranih komponent z UV/VIS spektroskopijo. Po analizi stabilnosti smo izvedli encimski test, kjer smo s pomočjo fluorogenega substrata merili intenziteto fluorescence. Iz dobljenih podatkov smo določili vrednosti EC50 preiskovanih komponent in ugotovili, da se inhibitorni efekt cinkovih soli poveča ob dodatku piritiona. Takšno znanje je uporabno predvsem pri načrtovanju eksperimentov, ki bi lahko pospešili tekmo iskanja potencialnih inhibitorjev encimov.

Keywords

cisteinski katepsini;katepsin L;encimska aktivnost;regulacija katepsinov;inhibitorji;pirition;cink;diplomska dela;

Data

Language: Slovenian
Year of publishing:
Typology: 2.11 - Undergraduate Thesis
Organization: UL FKKT - Faculty of Chemistry and Chemical Technology
Publisher: [R. Dajčman]
UDC: 577.15:546.47:547.82(043.2)
COBISS: 81768963 Link will open in a new window
Views: 274
Downloads: 54
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Other data

Secondary language: English
Secondary title: Effect of zinc salts and pyrithonal derivatives on cathepsine L activity
Secondary abstract: Enzymes are important targets in drug research to fight against pathogens and the body’s own disorders that lead to diseases states. At first, it seems that proteases do not belong to this group of enzymes, because they are only involved in protein turnover, which is completely wrong. One group of proteases; cysteine proteases, which are preferentially found in the lysosome, are also involved in many other cellular processes than just normal degradation of extracellular material and antigen processing. One of these enzymes is cathepsin L., which is involeved, among other things, in the development of severe acute respiratory syndrome (SARS). In the last decades, cysteine cathepsins have also become an interesting topic for studying the inhibitory effect. One of the most studied zinc complexes is zinc pyrithione, which has been long known for its antimicrobial properties and it is a key component in commercially available anti-dandruff shampoos. Beside antifungal activity, zinc pyrithione exhibits antiviral activity, making it a potential drug against viruses. Little is known about its effect on cathepsin L, but on the other hand, zinc ions are known to inhibit cathepsin B, which also belongs to CA1 family of papain-like proteases. For this reason, we have decided to examine the effect of pyrithione in combination with zinc salts on the cathepsin L activity. We first determined the stability of all tested components by UV/VIS spectroscopy. After that, we performed enzyme test, where the fluorescence intensity was measured using a fluorogenic substrate. From the obtained data, we determined the EC50 values of the tested components and found out that the inhibitory effect of zinc salts increases with the addition of pyrithione. Such knowledge is especially useful in designing an experiments, that would speed up the search for potential enzyme inhibitors.
Secondary keywords: cathepsin L;enzymatic activity;pyrithione;zinc;
Type (COBISS): Bachelor thesis/paper
Study programme: 1000371
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Fak. za kemijo in kemijsko tehnologijo, UNI Biokemija
Pages: 35 str.
ID: 13345766