Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing
Само за регистроване кориснике
2016
Аутори
Butorac, AnaMekic, Meliha Solak
Hozić, Amela
Diminic, Janko
Gamberger, Dragan
Nišavić, Marija
Cindrić, Mario
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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 signifi...cantly 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.
Извор:
Rapid Communications in Mass Spectrometry, 2016, 30, 14, 1687-1694Финансирање / пројекти:
- INNOMOL - Enhancement of the Innovation Potential in SEE through new Molecular Solutions in Research and Development (EU-FP7-316289)
- European Fund for Regional Development [STRIP/Aplimetafarma/RC.2.2.08-0046]
DOI: 10.1002/rcm.7594
ISSN: 0951-4198; 1097-0231
PubMed: 28328037
WoS: 000379975500003
Scopus: 2-s2.0-85016014909
Колекције
Институција/група
VinčaTY - 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 . .