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Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing
dc.creator | Butorac, Ana | |
dc.creator | Mekic, Meliha Solak | |
dc.creator | Hozić, Amela | |
dc.creator | Diminic, Janko | |
dc.creator | Gamberger, Dragan | |
dc.creator | Nišavić, Marija | |
dc.creator | Cindrić, Mario | |
dc.date.accessioned | 2018-03-01T17:03:19Z | |
dc.date.available | 2018-03-01T17:03:19Z | |
dc.date.issued | 2016 | |
dc.identifier.issn | 0951-4198 | |
dc.identifier.issn | 1097-0231 | |
dc.identifier.uri | https://vinar.vin.bg.ac.rs/handle/123456789/1183 | |
dc.description.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. | en |
dc.relation | info:eu-repo/grantAgreement/EC/FP7/316289/EU// | |
dc.relation | European Fund for Regional Development [STRIP/Aplimetafarma/RC.2.2.08-0046] | |
dc.rights | restrictedAccess | en |
dc.source | Rapid Communications in Mass Spectrometry | en |
dc.title | Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization de novo sequencing | en |
dc.type | article | en |
dc.rights.license | ARR | |
dcterms.abstract | Мекиц, Мелиха Солак; Нишавић Марија; Буторац, Aна; Гамбергер, Драган; Хозиц, Aмела; Циндриц, Марио; Диминиц, Јанко; | |
dc.citation.volume | 30 | |
dc.citation.issue | 14 | |
dc.citation.spage | 1687 | |
dc.citation.epage | 1694 | |
dc.identifier.wos | 000379975500003 | |
dc.identifier.doi | 10.1002/rcm.7594 | |
dc.citation.rank | M22 | |
dc.identifier.pmid | 28328037 | |
dc.type.version | publishedVersion | |
dc.type.version | publishedVersion | |
dc.identifier.scopus | 2-s2.0-85016014909 |