Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionization<i>de novo</i>sequencing

Butorac, Ana; Mekić, Meliha Solak; Hozić, Amela; Diminić, Janko; Gamberger, Dragan; Nišavić, Marija; Cindrić, Mario

doi:10.1002/rcm.7594

Cited by 5 publications

(9 citation statements)

References 28 publications

(52 reference statements)

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“…The algorithm utilizes b‐ions to derive a sequence as the FBSA derivatization enables the discriminatory detection of a predominantly single series of b‐ions in MS/MS positive‐ and negative mass spectra. The algorithm is described in detail by Butorac et al 12 Further, conversion of sulfonated b‐ions derived from negative tandem mass spectra into y‐ions is achieved in accordance with the spectrum transformation equation:

\frac{m}{z} {(y - ion)}^{+} goodbreak= \frac{m}{z} {(precursor - ion)}^{+} goodbreak- \frac{m}{z} {(Sulfo b - ion)}^{-} goodbreak- 1.00782 .

…”

Section: Methodsmentioning

confidence: 99%

“…Previous research based on peptide derivatization and de novo sequencing, using 5‐formyl‐1,3‐benzenedisulphonic acid (FBDSA) as a derivatization reagent, enabled de novo sequencing of Lactobacillus brevis derivate peptides in negative‐ and positive ion modes using matrix‐assisted laser desorption ionization‐ tandem mass spectrometry (MALDI‐MS/MS) 16 . Furthermore, a modified SPITC peptide derivatization procedure under acidic conditions (pH ≤5), which was used for peptide de novo sequencing of Fusarium delphinoides , produced signal intensities 6–10 times greater in comparison to commonly used SPITC derivatization under basic conditions 12 . Although the aforementioned derivatization reagents (FBDSA and SPITC) exhibited significant prospective for peptide‐directed fragmentation and de novo sequencing using MALDI techniques, both studies had disadvantages when analyzed using a UPLC‐nanoESI mass spectrometer under the given chromatographic conditions (optimal separation of underivatized bovine serum albumin [BSA] tryptic peptides).…”

Section: Introductionmentioning

confidence: 99%

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N‐Terminal chemical derivatization of peptides with 4‐formyl‐benzenesulfonic acid and electrospray positive and negative tandem mass spectrometry with high abundance of b‐ion series

Ozdanovac

Dončević

Hozić

et al. 2023

Rapid Comm Mass Spectrometry

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Selective derivatization of peptide N-terminus with 4-formylbenzenesulfonic acid (FBSA) enables chemically activated fragmentation in positive and negative ion modes (ESI+/À) under charge reduction conditions. Overlapped positive and negative tandem mass spectra show b-ions making the assignment of bion series fragments easy and accurate. Methods: We developed an FBSA-peptide microwave-assisted derivatization procedure. Derivatized and nonderivatized bovine serum albumin tryptic peptides and insulin non-tryptic peptide were compared after tandem mass spectrometry (MS/MS) analysis in positive and negative ion modes. A high-quality data set of sulfonated b-ions obtained in negative tandem mass spectra of singly charged FBSApeptides were matched to detected b-ions in positive MS/MS spectra. Moreover, negative spectra signals were converted and matched against y-ions in positive tandem mass spectra to identify complete peptide sequences. Results: The FBSA derivatization procedure produced a significantly improved MS/MS data set (populated by high-intensity signals of b-and y-ions) compared to commonly used N-terminal sulfonation reagents. Undesired side reactions almost do not occur, and the procedure reduces the derivatization time. It was found that b-ion intensities comprise 15% and 13% compared to combined ion intensities generated in positive-and negative ion modes, respectively. High visibility of b-ion series in negative ion mode can be attributed to N-terminal sulfonation that had no negative effect on the production of b-and y-ion series in positive ion mode. Conclusions:The FBSA derivatization and de novo sequencing approach outlined here is a reliable method for accurate peptide sequence assignment. Increased production of b-ions in positive-and negative ion modes greatly improves peak assignment and thus enables accurate sequence reconstruction. Implementation of the named methodology would improve the quality of de novo sequencing data and reduce the number of misinterpreted spectra.Luka Ozdanovac and Lucija Dončevi c are joint first authors.

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\frac{m}{z} {(y - ion)}^{+} goodbreak= \frac{m}{z} {(precursor - ion)}^{+} goodbreak- \frac{m}{z} {(Sulfo b - ion)}^{-} goodbreak- 1.00782 .

…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

N‐Terminal chemical derivatization of peptides with 4‐formyl‐benzenesulfonic acid and electrospray positive and negative tandem mass spectrometry with high abundance of b‐ion series

Ozdanovac

Dončević

Hozić

et al. 2023

Rapid Comm Mass Spectrometry

Self Cite

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“…Chemical derivatization changes the physicochemical properties of an analyte by introducing a modifying group into its certain functional groups, which can improve the detection sensitivity of the analyte, and exclude the matrix interference in samples 14–17 . Chemical derivatization increases the stability and the volatility of the sample, and improves the capacity of GC separation, which has been applied for the analysis of polar compounds by GC/mass spectrometry (MS) 18–21 .…”

Section: Introductionmentioning

confidence: 99%

Determination of trace fluoride in water samples by silylation and gas chromatography/mass spectrometry analysis

Chen

Wang

et al. 2021

Rapid Comm Mass Spectrometry

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Rationale Due to the ubiquity of fluorides and the small gap between a safe dose and a harmful one, it is necessary to develop a robust analytical method for determination of fluoride ions in various water samples with complex matrices. Methods Silylation of the fluoride ion was carried out by treatment with hydrochloric acid and phenyldimethylchlorosilane at room temperature. The formed phenyldimethylfluorosilane was detected by gas chromatography/mass spectrometry (GC/MS). Results Under the optimized conditions, linearity in the analytical method ranged from 0.050 to 5.0 μg/mL for the fluoride ion with R2 >0.9999. Reasonable reproducibility was obtained with the intraday relative standard deviation (RSD) (N = 5) of 2.04% and interday RSD (N = 5) of 3.75% at the concentration of 0.10 μg/mL. The developed method has been successfully applied to determine the fluoride ion in real water samples, including waste water samples, with the recovery between 81.12% and 113.04%. Conclusions A robust method for the determination of the fluoride ion has been developed by silylation with phenyldimethylchlorosilane and GC/MS analysis. The established method showed good anti‐interference and precision, and it has been applied for determination of the fluoride ion in various water samples.

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“…An efficient approach to enhance fragmentation efficiency of nontryptic peptides is to introduce positively charged basic groups by means of chemical derivatization. [15][16][17][18] Over the past decades, plenty of chemical derivatization strategies based on amino, 19,20 and carboxyl groups [21][22][23] had been utilized to facilitate peptide fragmentation. Carboxy groups derivatization offers a compelling approach for enhancing charge states and peptide fragmentation because these sites are typically among the most acidic in peptides and thus most susceptible to deprotonation.…”

Section: Introductionmentioning

confidence: 99%

“…However, in the aforementioned alternative digestion modes, the lack of C‐terminal basic amino acids of these nontryptic peptides would not benefit for peptide fragmentation. An efficient approach to enhance fragmentation efficiency of nontryptic peptides is to introduce positively charged basic groups by means of chemical derivatization 15–18 …”

Section: Introductionmentioning

confidence: 99%

Sequential amidation of peptide C‐termini for improving fragmentation efficiency

Tian

Yang

et al. 2020

J Mass Spectrom

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Owing to the poor fragmentation efficiency caused by the lack of a positively charged basic group at the C-termini of peptides, the identification of nontryptic peptides in classical proteomics is known to be less efficient. Particularly, attaching positively charged basic groups to C-termini via chemical derivatizations is known to be able to enhance their fragmentation efficiency. In this study, we introduced a novel strategy, C-termini sequential amidation reaction (CSAR), to improve peptide fragmentation efficiency. By this strategy, C-terminal and side-chain carboxyl groups were firstly amidated by neutral methylamine (MA), and then C-terminal amide bonds were selectively deamidated through peptide amidase while side-chain amide bonds remained unchanged, followed by the secondary amidation of C-termini via basic agmatine (AG). We optimized the amidation reaction conditions to achieve the MA derivatization efficiency of >99% for side-chain carboxyl groups and AG derivatization efficiency of 80% for the hydrolytic C-termini. We applied CSAR strategy to identify bovine serum albumin (BSA) chymotryptic digests, resulting in the increased fragmentation efficiencies (improvement by 9-32%) and charge states (improvement by 39-52%) under single or multiple dissociation modes. The strategy described here might be a promising approach for the identification of peptides that suffered from poor fragmentation efficiency.

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Benefits of selective peptide derivatization with sulfonating reagent at acidic pH for facile matrix-assisted laser desorption/ionizationde novosequencing

Cited by 5 publications

References 28 publications

N‐Terminal chemical derivatization of peptides with 4‐formyl‐benzenesulfonic acid and electrospray positive and negative tandem mass spectrometry with high abundance of b‐ion series

N‐Terminal chemical derivatization of peptides with 4‐formyl‐benzenesulfonic acid and electrospray positive and negative tandem mass spectrometry with high abundance of b‐ion series

Determination of trace fluoride in water samples by silylation and gas chromatography/mass spectrometry analysis

Sequential amidation of peptide C‐termini for improving fragmentation efficiency

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