prikaz prve stranice dokumenta Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )
Embargo period 2027-04-03
master's thesis
Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )
2024. urn:nbn:hr:166:852531
Blajić, Ante
University of Split
Faculty of Science

Cite this document

Blajić, A. (2024). Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. ) (Master's thesis). Split: University of Split, Faculty of Science. Retrieved from https://urn.nsk.hr/urn:nbn:hr:166:852531

Blajić, Ante. "Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )." Master's thesis, University of Split, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:166:852531

Blajić, Ante. "Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )." Master's thesis, University of Split, Faculty of Science, 2024. https://urn.nsk.hr/urn:nbn:hr:166:852531

Blajić, A. (2024). 'Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )', Master's thesis, University of Split, Faculty of Science, accessed 08 December 2024, https://urn.nsk.hr/urn:nbn:hr:166:852531

Blajić A. Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. ) [Master's thesis]. Split: University of Split, Faculty of Science; 2024 [cited 2024 December 08] Available at: https://urn.nsk.hr/urn:nbn:hr:166:852531

A. Blajić, "Optimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )", Master's thesis, University of Split, Faculty of Science, Split, 2024. Available at: https://urn.nsk.hr/urn:nbn:hr:166:852531

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Metadata
TitleOptimizacija metoda ekstrakcije proteina za LC-MS/MS analizu: Preliminarna analiza proteoma biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten. )
Title (english)Optimization of protein extraction methods for LC-MS/MS analysis: Preliminary analysis of the proteome of the Croatian oak plant (Quercus virgiliana (Ten.) Ten. )
AuthorAnte Blajić
MentorIvica Šamanić (mentor)
MentorStjepan Orhanović (komentor)
MentorMarija Nazlić (pomoć pri izradi, neposredni voditelj)
Committee memberStjepan Orhanović (predsjednik povjerenstva)
Committee memberIvica Šamanić (član povjerenstva)
Committee memberMarija Nazlić (član povjerenstva)
GranterUniversity of Split
Faculty of Science
Split
Defense date and country2024-09-27, Croatia
Scientific / art field,
discipline and subdiscipline		NATURAL SCIENCES
Biology
Abstract
U ovome radu proveli smo analizu metoda ekstrakcije proteina za opisivanje proteoma lista i tkiva modificiranog hrastovom osicom biljke hrvatskog hrasta (drmun, Quercus virgiliana (Ten.) Ten.) pomoću LC-MS spektrometrije masa. Testirana su četiri različita protokola kako bi se odredila najučinkovitija metoda za ekstrakciju i izolaciju proteoma: SDC-FASP, ekstrakcija fenolom u kombinaciji sa SDC-FASP, taloženje TCA/acetonom praćeno ekstrakcijom fenolom i SDC-FASP, te taloženje TCA/acetonom u kombinaciji sa SDC-FASP. Cilj je bio utvrditi najučinkovitiju metodu za izolaciju i pročišćavanje proteina za pripremu uzoraka uz pomoć filtera (engl. filter-assisted sample preparation, FASP). Identifikacija proteina i posttranslacijske modifikacije (PTM) analizirane su korištenjem softvera ProteinPilot™ prema bazi podataka referentnog proteoma Quercus lobata. Iako su izmjereni niski prinosi proteina za sve protokole, taloženje TCA/acetonom praćeno ekstrakcijom fenolom i SDC-FASP dalo je najveći broj identificiranih proteina. Ukupni ionski kromatogrami (engl. total ion chromatograms, TIC) također su potvrdili niske signale peptidnih iona koji ukazuju na niske koncentracije proteina. Usporedna analiza uzoraka lista i tkiva modificiranog hrastovom osicom (engl. gall-wasp) pokazala je različite proteomske profile. Tkiva lista bila su bogata proteinima povezanim s fotosintezom, energetskim metabolizmom i odgovorima na stres, dok tkiva modificiranog hrastovom osicom sadrže proteine povezane s prikupljanjem hranjivih tvari, regulacijom razvoja i imunološkom modulacijom, odražavajući fiziološke učinke modificiranog tkiva na domaćina.
Abstract (english)
In this study, we performed an analysis of protein extraction methods suitable for proteomic experiments of oak leaves and oak gall wasp-modified tissue from Quercus virgiliana using LC-MS-based mass spectrometry. Four extraction protocols were tested for optimal protein isolation: SDC-FASP, phenol extraction combined with SDC-FASP, TCA/acetone precipitation with phenol extraction and SDC-FASP, and a combination of TCA/acetone precipitation and SDC-FASP. The aim was to determine the most effective method for protein isolation and purification for filter-assisted sample preparation (FASP). Protein identification and post-translational modifications (PTMs) were analyzed using ProteinPilot™ with reference to the Quercus lobata proteome. While low protein yields were observed for all protocols, TCA/acetone precipitation followed by phenol extraction and SDC-FASP yielded the highest number of identified proteins. The total ion chromatograms (TIC) also confirmed the low peptide ion signals indicating low protein concentrations. Comparative analysis of leaf and gall wasp-modified tissues revealed different proteomic profiles. The leaf tissues were rich in proteins related to photosynthesis, energy metabolism and stress responses, while gall wasp-modified tissues had proteins related to nutrient acquisition, developmental regulation and immune modulation, reflecting the physiological effects of gall production on the host.
Keywords
Keywords (english)
Languagecroatian
URN:NBNurn:nbn:hr:166:852531
Study programmeTitle: Biology and Chemistry; specializations in: specialization in Education
Course: specialization in Education
Study programme type: university
Study level: graduate
Academic / professional title: sveučilišni/a magistar/magistra edukacije biologije i kemije (sveučilišni/a magistar/magistra edukacije biologije i kemije)
Type of resourceText
File originBorn digital
Access conditionsEmbargoed access
Embargo expiration date: 2027-04-03
Terms of useLicence In copyright icon
Created on2024-10-24 12:32:01