Naslov
Derivatisation of linaclotide and bovine serum albumin with acrylamide and 4-vinylpiridine analyzded by MALDI-TOF/TOF and nanoUPLC coupled to the ESI-QTOF

Autori
Ozdanovac, Luka ; Dončević, Lucija ; Cindrić, Mario

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
6. simpozij studenata doktorskih studija PMF-a : knjiga sažetaka = 6th Faculty of Science PhD student symposium : book of abstracts / Schneider, Petra - Zagreb : Prirodoslovno-matematički fakultet Sveučilišta u Zagrebu, 2022, 78-79

ISBN
978-953-6076-93-2

Skup
6. Simpozij studenata doktorskih studija PMF-a = 6th Faculty of Science PhD Student Symposium

Mjesto i datum
Zagreb, Hrvatska, 23.04.2022. - 24.04.2022

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Domaća recenzija

Ključne riječi
Protein derivatization ; Selective alkylation ; MALDI-TOF/TOF ; nanoUPLC-ESI-QTOF ; Proteomics identification

Sažetak
Proteomics identification and quantification techniques often involve the process of protein derivatization. Binding of different ions or molecules to specific sites of the protein is used to modify structural, ionic, hydrophobic-hydrophilic, and other protein properties. Changes in structural properties allow better chromatographic or ionic separation of protein or protein fragments. At the end, derivatization is carried out to increase overall sensitivity of the analysis. To improve and accelerate protein derivatization procedures of cysteine branches of peptides or proteins, two derivatization reagents, acrylamide and 4- vinylpyridine, were applied [1]. Reactions were optimized using Linaclotide peptide that contains six cysteines interconnected with three disulfide bridges (Figure 1. A). Prior to mass spectrometric analysis derivatization procedure was conducted on bovine serum albumin protein followed by qualitative and quantitative analysis of derivatized tryptic fragments (Figure 1. B). Selective alkylation of the cysteine side branches was achieved after reduction of disulfide bridges. In order to speed up alkylation reaction microwave irradiation was utilized for 8 minutes with radiation power of 180 W. The analysis of peptide reaction mixtures was performed using MALDI-TOF/TOF and nanoUPLC-ESI- QTOF mass spectrometers. Collected spectra were processed with the software search package ProteinLynx Global SERVERTM (PLGS).

Izvorni jezik
Engleski

Znanstvena područja
Kemija

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