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Načrtovanje, izražanje in ovrednotenje heterodimernih obvitih vijačnic na osnovi tropomiozina = Design, production and characterization of tropomyosin based coiled-coil dimersMežnaršič, EvaDobro poznavanje pravil zvijanja različnih naravnih proteinov nam omogoča načrtovanje proteinskih nanostruktur, kakršnih ne najdemo v naravi, de novo. Nove strukture lahko uporabimo v medicini, ... kemiji in biotehnologiji, npr. za ciljano dostavo zdravil v tkiva, razvoj cepiv, tkivno inženirstvo, načrtovanje biosenzorjev, biokatalizo in razvoj bionanomaterialov. Za načrtovanje de novo dovolj dobro poznamo le nekaj manjših naravnih domen proteinov, ki jih lahko uporabimo kot module za sestavljanje večjih poliedrskih nanostruktur. Proteinski origami je primer modularne topološko pogojene nanostrukture, sestavljene iz obvitih vijačnic, ki predstavljajo module. Posamezni moduli interagirajo izključno s svojim načrtovanim partnerjem in se tako zvijejo v načrtovano strukturo. Obvite vijačnice so polipeptidna zvitja dveh verig, za katera je značilen vzorec sedmih aminokislinskih ostankov, imenovanih heptade. V sklopu magistrskega dela smo načrtovali, izrazili in ovrednotili tri obvite vijačnice: Tropo-Cn6/Tropo-Dn6, Tropo-Cn12/Tropo-Dn12 in Tropo-Cn14/Tropo-Dn14. Te so del pete generacije heterodimernih obvitih vijačnic, dolgih 15 heptad, na osnovi tropomiozina, načrtovanih na Kemijskem inštitutu. V peti generaciji smo izbrali najbolj obetaven par iz četrte generacije in uvedli spremembe v devet prej nemodificiranih heptad. Velikost načrtovanih polipeptidov smo določili z metodo dinamičnega sipanja svetlobe (DLS) in ugotovili, da se je skladala s predvideno velikostjo okoli 15 nm. Delež sekundarne vijačne strukture smo ugotavljali z metodo cirkularnega dikroizma (CD). Pri paru Tropo-Cn6/Tropo-Dn6 v nasprotju s pričakovanji nismo opazili vijačnih struktur. Para Tropo-Cn12/Tropo-Dn12 in Tropo-Cn14/Tropo-Dn14 sta dala boljše rezultate. Z merjenjem temperaturne stabilnosti sekundarnih vijačnih struktur smo ugotovili, da je pri paru Tropo-Cn12/Tropo-Dn12 večja razlika med Tm posameznega polipeptida in Tm para. Glede na rezultate smo se zato odločili, da bomo nadaljnjo optimizacijo heterodimernih obvitih vijačnic na osnovi tropomiozina nadaljevali na podlagi Tropo-Cn12/Tropo-Dn12.Type of material - master's thesis ; adult, seriousPublication and manufacture - Ljubljana : [E. Mežnaršič], 2019Language - slovenianCOBISS.SI-ID - 4825713
Author
Mežnaršič, Eva
Other authors
Bratkovič, Tomaž, farmacevt, 1978- |
Gradišar, Helena
Topics
Beljakovine |
Tkivno inženirstvo |
obvita vijačnica |
tropomiozin |
proteinski origami |
načrtovanje de novo |
polipeptidi
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Library/institution |
City | Acronym | For loan | Other holdings |
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Faculty of Pharmacy, Ljubljana | Ljubljana | FFALJ |
reading room 1 cop.
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Database name | Field | Year |
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Links to authors' personal bibliographies | Links to information on researchers in the SICRIS system |
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Mežnaršič, Eva | ![]() |
Bratkovič, Tomaž, farmacevt, 1978- | 23598 |
Gradišar, Helena | 17915 |
Source: Personal bibliographies
and: SICRIS
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