TY  - JOUR
AU  - Butorac, Ana
AU  - Mekic, Meliha Solak
AU  - Hozić, Amela
AU  - Diminic, Janko
AU  - Gamberger, Dragan
AU  - Nišavić, Marija
AU  - Cindrić, Mario
PY  - 2016
UR  - https://vinar.vin.bg.ac.rs/handle/123456789/1183
AB  - RATIONALE: One of the most challenging tasks of proteomics is peptide de novo sequencing. 4-Sulfophenyl isothiocyanate (SPITC) peptide derivatization enables acquisition of high-quality tandem mass spectra (MS/MS) for de novo sequencing, but unwanted non-specific reactions and reduced mass spectra (MS) signal intensities still represent the obstacles in highthroughput de novo sequencing. METHODS: We developed a SPITC peptide derivatization procedure under acidic conditions (pH LT = 5). Derivatized peptides were analyzed by matrix-assisted laser desorption/ionization (MALDI-MS) in negative ion mode followed by MS/MS in positive ion mode. A de novo sequencing tool, named DUST, adjusted to SPITC chemistry, was designed for successful high-throughput peptide de novo sequencing. This high-throughput peptide de novo sequencing was tested on Fusarium delphinoides, an organism with an uncharacterized genome. RESULTS: The SPITC derivatization procedure under acidic conditions produced a significantly improved MS dataset in comparison to commonly used derivatization under basic conditions. Signal intensities were 6 to 10 times greater and the over-sulfonation effect measured on lysine-containing peptides was significantly decreased. Furthermore, development of a novel DUST algorithm enabled automated de novo sequencing with the calculated accuracy of 70.6%. CONCLUSIONS: The SPITC derivatization and de novo sequencing approach outlined here provides a reliable method for high-throughput peptide de novo sequencing. High-throughput peptide de novo sequencing enabled protein mutation identification and identification of proteins from organisms with non-sequenced genomes. Copyright (C) 2016 John Wiley and Sons, Ltd.
T2  - Rapid Communications in Mass Spectrometry
T1  - Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing
VL  - 30
IS  - 14
SP  - 1687
EP  - 1694
DO  - 10.1002/rcm.7594
ER  - 

@article{
author = "Butorac, Ana and Mekic, Meliha Solak and Hozić, Amela and Diminic, Janko and Gamberger, Dragan and Nišavić, Marija and Cindrić, Mario",
year = "2016",
abstract = "RATIONALE: One of the most challenging tasks of proteomics is peptide de novo sequencing. 4-Sulfophenyl isothiocyanate (SPITC) peptide derivatization enables acquisition of high-quality tandem mass spectra (MS/MS) for de novo sequencing, but unwanted non-specific reactions and reduced mass spectra (MS) signal intensities still represent the obstacles in highthroughput de novo sequencing. METHODS: We developed a SPITC peptide derivatization procedure under acidic conditions (pH LT = 5). Derivatized peptides were analyzed by matrix-assisted laser desorption/ionization (MALDI-MS) in negative ion mode followed by MS/MS in positive ion mode. A de novo sequencing tool, named DUST, adjusted to SPITC chemistry, was designed for successful high-throughput peptide de novo sequencing. This high-throughput peptide de novo sequencing was tested on Fusarium delphinoides, an organism with an uncharacterized genome. RESULTS: The SPITC derivatization procedure under acidic conditions produced a significantly improved MS dataset in comparison to commonly used derivatization under basic conditions. Signal intensities were 6 to 10 times greater and the over-sulfonation effect measured on lysine-containing peptides was significantly decreased. Furthermore, development of a novel DUST algorithm enabled automated de novo sequencing with the calculated accuracy of 70.6%. CONCLUSIONS: The SPITC derivatization and de novo sequencing approach outlined here provides a reliable method for high-throughput peptide de novo sequencing. High-throughput peptide de novo sequencing enabled protein mutation identification and identification of proteins from organisms with non-sequenced genomes. Copyright (C) 2016 John Wiley and Sons, Ltd.",
journal = "Rapid Communications in Mass Spectrometry",
title = "Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing",
volume = "30",
number = "14",
pages = "1687-1694",
doi = "10.1002/rcm.7594"
}

Butorac, A., Mekic, M. S., Hozić, A., Diminic, J., Gamberger, D., Nišavić, M.,& Cindrić, M.. (2016). Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing. in Rapid Communications in Mass Spectrometry, 30(14), 1687-1694.
https://doi.org/10.1002/rcm.7594

Butorac A, Mekic MS, Hozić A, Diminic J, Gamberger D, Nišavić M, Cindrić M. Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing. in Rapid Communications in Mass Spectrometry. 2016;30(14):1687-1694.
doi:10.1002/rcm.7594 .

Butorac, Ana, Mekic, Meliha Solak, Hozić, Amela, Diminic, Janko, Gamberger, Dragan, Nišavić, Marija, Cindrić, Mario, "Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing" in Rapid Communications in Mass Spectrometry, 30, no. 14 (2016):1687-1694,
https://doi.org/10.1002/rcm.7594 . .