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master's thesis
Mapping of sufrace-exposed cysteines in ATPase ISWI
Zagreb: University of Zagreb, Faculty of Science, 2017. urn:nbn:hr:217:949570
Vizjak, Petra
University of Zagreb
Faculty of Science
Department of Chemistry

Institutional repository: Repository of the Faculty of Science

Cite this document

Vizjak, P. (2017). Mapping of sufrace-exposed cysteines in ATPase ISWI (Master's thesis). Zagreb: University of Zagreb, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:217:949570

Vizjak, Petra. "Mapping of sufrace-exposed cysteines in ATPase ISWI." Master's thesis, University of Zagreb, Faculty of Science, 2017. https://urn.nsk.hr/urn:nbn:hr:217:949570

Vizjak, Petra. "Mapping of sufrace-exposed cysteines in ATPase ISWI." Master's thesis, University of Zagreb, Faculty of Science, 2017. https://urn.nsk.hr/urn:nbn:hr:217:949570

Vizjak, P. (2017). 'Mapping of sufrace-exposed cysteines in ATPase ISWI', Master's thesis, University of Zagreb, Faculty of Science, accessed 22 November 2024, https://urn.nsk.hr/urn:nbn:hr:217:949570

Vizjak P. Mapping of sufrace-exposed cysteines in ATPase ISWI [Master's thesis]. Zagreb: University of Zagreb, Faculty of Science; 2017 [cited 2024 November 22] Available at: https://urn.nsk.hr/urn:nbn:hr:217:949570

P. Vizjak, "Mapping of sufrace-exposed cysteines in ATPase ISWI", Master's thesis, University of Zagreb, Faculty of Science, Zagreb, 2017. Available at: https://urn.nsk.hr/urn:nbn:hr:217:949570

Metadata
TitleMapping of sufrace-exposed cysteines in ATPase ISWI
Title (croatian)Mapiranje površinski izloženih cisteina u ATP-azi ISWI
AuthorPetra Vizjak
MentorFelix Müller-Planitz (mentor)
Committee memberMarko Močibob (predsjednik povjerenstva)
Committee memberĐani Škalamera (član povjerenstva)
Committee memberBranimir Bertoša (član povjerenstva)
Committee memberJasmina Rokov Plavec (član povjerenstva)
GranterUniversity of Zagreb
Faculty of Science
(Department of Chemistry)
Zagreb
Defense date and country2017-09-29, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Chemistry
Abstract
In an eukaryotic cell, DNA molecule exists in the form of chromatin. The repetitive unit of chromatin is a nucleosome, 147 bp of DNA wrapped around histone octamer. Nucleosome position along DNA is important for transcription regulation. Chromatin remodeler ISWI (Drosophila melanogaster) slides nucleosomes along DNA. The aim of this work is to map surface-exposed cysteines in the protein ISWI. The surface-exposed cysteines are easily avaliable for chemical reaction. To probe accessibility of cysteines for chemical reaction, commonly used reagents for spectrophotometric detection of thiols were used: 5,5ʹ-dithiobis- (2-nitrobenzoic acid) and 2-nitro-5-thiocyanatobenzoic acid. To unequivocally identify surfaceexposed cysteines, ISWI was treated with primary alkylating reagent (N-ethylmaleimide), denaturated and treated with secondary alkylating reagent (iodoacetic acid). The two reactions add to the cysteine different modification groups which can be discriminated by mass spectrometry. Half of the total cysteines could be clasified either as buried or surface-exposed in the protein structure.
Abstract (croatian)
U eukariotskoj stanici, molekula DNA nalazi se u obliku kromatina. Ponavljajuća jedinica kromatina je nukleosom, 147 parova baza DNA namotano oko histonskog oktamera. Položaj nukleosoma na DNA ima važnu ulogu u kontroli transkripcije. Remodelirajući protein ISWI (Drosophila melanogaster) pomiče nukleosome duž DNA. Cilj ovog rada je mapirati površinski izložene cisteine proteina ISWI. Rezultati će kasnije biti iskorišteni za analizu postojećih strukturnih modela proteina ISWI. Cisteini koji se nalaze na površini proteina, brže podliježu kemijskim reakcijama. Kako bi se potvrdilo da cisteini reagiraju različitom kinetikom, korišteni su reagensi za spektrofotometrijsko utvrđivanje tiola: 5,5ʹ-ditiobis-(2- nitrobenzojeva kiselina) i 2-nitro-5-tiocianatobenzojeva kiselina. Za točnu identifikaciju površinski izloženih cisteina, ISWI je tretiran s primarnim alkilirajućim reagensom (Netilmaleimidom), denaturiran i tretiran sa sekundarnim alkilirajućim reagensom (jodoctenom kiselinom). Produkt tih dviju reakcija je dodatak različitih modifikacijskih skupina na cisteine, koje se potom mogu razlučiti spektrometrijom masa. Polovica od ukupnih cisteina mogla se klasificirati kao površinski izloženi ili zaklonjeni unutar strukture proteina.
Keywords
Keywords (croatian)
Languageenglish
URN:NBNurn:nbn:hr:217:949570
Study programmeTitle: Graduate university studies in chemistry; specializations in: Research programme
Course: Research programme
Study programme type: university
Study level: graduate
Academic / professional title: magistar/magistra kemije (magistar/magistra kemije)
Type of resourceText
File originBorn digital
Access conditionsOpen access
Terms of useLicence In copyright icon
Created on2017-10-20 10:51:25