Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells
Mass spectrometry methods are commonly used in the identification of peptides and biomarkers. Due to a relatively low abundance of proteins in biological samples, there is a need for the development of novel derivatization methods that would improve MS detection limits. Hence, novel fluorescent <...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/21/24/9643 |
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| author | Joanna Fedorowicz Magdalena Wierzbicka Marek Cebrat Paulina Wiśniewska Rafał Piątek Beata Zalewska-Piątek Zbigniew Szewczuk Jarosław Sączewski |
| author_facet | Joanna Fedorowicz Magdalena Wierzbicka Marek Cebrat Paulina Wiśniewska Rafał Piątek Beata Zalewska-Piątek Zbigniew Szewczuk Jarosław Sączewski |
| author_sort | Joanna Fedorowicz |
| collection | DOAJ |
| description | Mass spectrometry methods are commonly used in the identification of peptides and biomarkers. Due to a relatively low abundance of proteins in biological samples, there is a need for the development of novel derivatization methods that would improve MS detection limits. Hence, novel fluorescent <i>N</i>–hydroxysuccinimide esters of dihydro-[1,2,4]triazolo[4,3-<i>a</i>]pyridin-2-ium carboxylates (<i>Safirinium P</i> dyes) have been synthesized. The obtained compounds, which incorporate quaternary ammonium salt moieties, easily react with aliphatic amine groups of peptides, both in solution and on the solid support; thus, they can be applied for derivatization as ionization enhancers. <i>Safirinium</i> tagging experiments with ubiquitin hydrolysate revealed that the sequence coverage level was high (ca. 80%), and intensities of signals were enhanced up to 8-fold, which proves the applicability of the proposed tags in the bottom–up approach. The obtained results confirmed that the novel compounds enable the detection of trace amounts of peptides, and fixed positive charge within the tags results in high ionization efficiency. Moreover, <i>Safirinium</i> NHS esters have been utilized as imaging agents for fluorescent labeling and the microscopic visualization of living cells such as <i>E. coli</i> Top10 bacterial strain. |
| first_indexed | 2024-03-10T13:59:08Z |
| format | Article |
| id | doaj.art-72d9147a6eb443d88dc589a83814b7a8 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T13:59:08Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-72d9147a6eb443d88dc589a83814b7a82023-11-21T01:22:11ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-12-012124964310.3390/ijms21249643Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial CellsJoanna Fedorowicz0Magdalena Wierzbicka1Marek Cebrat2Paulina Wiśniewska3Rafał Piątek4Beata Zalewska-Piątek5Zbigniew Szewczuk6Jarosław Sączewski7Department of Chemical Technology of Drugs, Faculty of Pharmacy, Medical University of Gdańsk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, PolandFaculty of Chemistry, University of Wrocław, ul. F. Joliot-Curie 14, 50-383 Wrocław, PolandFaculty of Chemistry, University of Wrocław, ul. F. Joliot-Curie 14, 50-383 Wrocław, PolandDepartment of Organic Chemistry, Faculty of Pharmacy, Medical University of Gdańsk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, PolandDepartment of Molecular Biotechnology and Microbiology, Chemical Faculty, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, PolandDepartment of Molecular Biotechnology and Microbiology, Chemical Faculty, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, PolandFaculty of Chemistry, University of Wrocław, ul. F. Joliot-Curie 14, 50-383 Wrocław, PolandDepartment of Organic Chemistry, Faculty of Pharmacy, Medical University of Gdańsk, Al. Gen. J. Hallera 107, 80-416 Gdańsk, PolandMass spectrometry methods are commonly used in the identification of peptides and biomarkers. Due to a relatively low abundance of proteins in biological samples, there is a need for the development of novel derivatization methods that would improve MS detection limits. Hence, novel fluorescent <i>N</i>–hydroxysuccinimide esters of dihydro-[1,2,4]triazolo[4,3-<i>a</i>]pyridin-2-ium carboxylates (<i>Safirinium P</i> dyes) have been synthesized. The obtained compounds, which incorporate quaternary ammonium salt moieties, easily react with aliphatic amine groups of peptides, both in solution and on the solid support; thus, they can be applied for derivatization as ionization enhancers. <i>Safirinium</i> tagging experiments with ubiquitin hydrolysate revealed that the sequence coverage level was high (ca. 80%), and intensities of signals were enhanced up to 8-fold, which proves the applicability of the proposed tags in the bottom–up approach. The obtained results confirmed that the novel compounds enable the detection of trace amounts of peptides, and fixed positive charge within the tags results in high ionization efficiency. Moreover, <i>Safirinium</i> NHS esters have been utilized as imaging agents for fluorescent labeling and the microscopic visualization of living cells such as <i>E. coli</i> Top10 bacterial strain.https://www.mdpi.com/1422-0067/21/24/9643fluorescence microscopyfluorescent probeH/D exchangelive cell imagingionization tagmass spectrometry |
| spellingShingle | Joanna Fedorowicz Magdalena Wierzbicka Marek Cebrat Paulina Wiśniewska Rafał Piątek Beata Zalewska-Piątek Zbigniew Szewczuk Jarosław Sączewski Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells International Journal of Molecular Sciences fluorescence microscopy fluorescent probe H/D exchange live cell imaging ionization tag mass spectrometry |
| title | Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells |
| title_full | Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells |
| title_fullStr | Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells |
| title_full_unstemmed | Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells |
| title_short | Application of <i>Safirinium N</i>-Hydroxysuccinimide Esters to Derivatization of Peptides for High-Resolution Mass Spectrometry, Tandem Mass Spectrometry, and Fluorescent Labeling of Bacterial Cells |
| title_sort | application of i safirinium n i hydroxysuccinimide esters to derivatization of peptides for high resolution mass spectrometry tandem mass spectrometry and fluorescent labeling of bacterial cells |
| topic | fluorescence microscopy fluorescent probe H/D exchange live cell imaging ionization tag mass spectrometry |
| url | https://www.mdpi.com/1422-0067/21/24/9643 |
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