Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time
- Autores
- Yu, Zhan; Chen, Lee Chuin; Mandal, Mridul Kanti; Nonami, Hiroshi; Erra Balsells, Rosa; Hiraoka, Kenzo
- Año de publicación
- 2012
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Although there are a lot of well established methods for monitoring enzyme-catalyzed reactions, most of them are based on changes in spectroscopic properties during the conversion of substrates to products. However, reactions without optical changes are common, which are inapplicable to these spectroscopic methods. As an alternative technique for enzymologic research, mass spectrometry (MS) is favored due to its specificity, sensitivity, and the ability to obtain stoichiometric information. In this work, probe electrospray ionization (PESI) source coupled with a time of flight mass spectrometer was employed to monitor some typical proteasecatalyzed reactions, including pepsinolysis and trypsinolysis of cytochrome c in real time. Due to the high electrical conductivity of each reaction system, corona discharges are likely to occur, which would decrease intensities of mass spectrometric signals. An ultra-fine sampling probe and an auxiliary vapor spray were adopted to prevent corona discharges. Experimental results from peptic and tryptic digestions of cytochrome c showed different and characteristic catalytic pathways. With the data presented in this study, PESI-MS can be considered as a potential tool for real-time monitoring of enzymatic reactions because of its simplicity in instrumental configuration, wide applicability under harsh conditions, and flexibility in combination with other techniques.
Fil: Yu, Zhan. Shenyang Normal University; China. University Of Yamanashi; Japón
Fil: Chen, Lee Chuin. University Of Yamanashi; Japón
Fil: Mandal, Mridul Kanti. University Of Yamanashi; Japón
Fil: Nonami, Hiroshi. Ehime University; Japón
Fil: Erra Balsells, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; Argentina
Fil: Hiraoka, Kenzo. University Of Yamanashi; Japón - Materia
-
Electrochemical Etching
Onlinemonitoring
Probe Electrospray Ionization
Proteolytic Reactions
Solid Probe - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/68168
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Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real timeYu, ZhanChen, Lee ChuinMandal, Mridul KantiNonami, HiroshiErra Balsells, RosaHiraoka, KenzoElectrochemical EtchingOnlinemonitoringProbe Electrospray IonizationProteolytic ReactionsSolid Probehttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Although there are a lot of well established methods for monitoring enzyme-catalyzed reactions, most of them are based on changes in spectroscopic properties during the conversion of substrates to products. However, reactions without optical changes are common, which are inapplicable to these spectroscopic methods. As an alternative technique for enzymologic research, mass spectrometry (MS) is favored due to its specificity, sensitivity, and the ability to obtain stoichiometric information. In this work, probe electrospray ionization (PESI) source coupled with a time of flight mass spectrometer was employed to monitor some typical proteasecatalyzed reactions, including pepsinolysis and trypsinolysis of cytochrome c in real time. Due to the high electrical conductivity of each reaction system, corona discharges are likely to occur, which would decrease intensities of mass spectrometric signals. An ultra-fine sampling probe and an auxiliary vapor spray were adopted to prevent corona discharges. Experimental results from peptic and tryptic digestions of cytochrome c showed different and characteristic catalytic pathways. With the data presented in this study, PESI-MS can be considered as a potential tool for real-time monitoring of enzymatic reactions because of its simplicity in instrumental configuration, wide applicability under harsh conditions, and flexibility in combination with other techniques.Fil: Yu, Zhan. Shenyang Normal University; China. University Of Yamanashi; JapónFil: Chen, Lee Chuin. University Of Yamanashi; JapónFil: Mandal, Mridul Kanti. University Of Yamanashi; JapónFil: Nonami, Hiroshi. Ehime University; JapónFil: Erra Balsells, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; ArgentinaFil: Hiraoka, Kenzo. University Of Yamanashi; JapónSpringer2012-04info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/68168Yu, Zhan; Chen, Lee Chuin; Mandal, Mridul Kanti; Nonami, Hiroshi; Erra Balsells, Rosa; et al.; Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time; Springer; Journal of The American Society for Mass Spectrometry; 23; 4; 4-2012; 728-7351044-0305CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1007/s13361-011-0323-5info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs13361-011-0323-5info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:01:05Zoai:ri.conicet.gov.ar:11336/68168instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:01:06.22CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
title |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
spellingShingle |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time Yu, Zhan Electrochemical Etching Onlinemonitoring Probe Electrospray Ionization Proteolytic Reactions Solid Probe |
title_short |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
title_full |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
title_fullStr |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
title_full_unstemmed |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
title_sort |
Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time |
dc.creator.none.fl_str_mv |
Yu, Zhan Chen, Lee Chuin Mandal, Mridul Kanti Nonami, Hiroshi Erra Balsells, Rosa Hiraoka, Kenzo |
author |
Yu, Zhan |
author_facet |
Yu, Zhan Chen, Lee Chuin Mandal, Mridul Kanti Nonami, Hiroshi Erra Balsells, Rosa Hiraoka, Kenzo |
author_role |
author |
author2 |
Chen, Lee Chuin Mandal, Mridul Kanti Nonami, Hiroshi Erra Balsells, Rosa Hiraoka, Kenzo |
author2_role |
author author author author author |
dc.subject.none.fl_str_mv |
Electrochemical Etching Onlinemonitoring Probe Electrospray Ionization Proteolytic Reactions Solid Probe |
topic |
Electrochemical Etching Onlinemonitoring Probe Electrospray Ionization Proteolytic Reactions Solid Probe |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.4 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Although there are a lot of well established methods for monitoring enzyme-catalyzed reactions, most of them are based on changes in spectroscopic properties during the conversion of substrates to products. However, reactions without optical changes are common, which are inapplicable to these spectroscopic methods. As an alternative technique for enzymologic research, mass spectrometry (MS) is favored due to its specificity, sensitivity, and the ability to obtain stoichiometric information. In this work, probe electrospray ionization (PESI) source coupled with a time of flight mass spectrometer was employed to monitor some typical proteasecatalyzed reactions, including pepsinolysis and trypsinolysis of cytochrome c in real time. Due to the high electrical conductivity of each reaction system, corona discharges are likely to occur, which would decrease intensities of mass spectrometric signals. An ultra-fine sampling probe and an auxiliary vapor spray were adopted to prevent corona discharges. Experimental results from peptic and tryptic digestions of cytochrome c showed different and characteristic catalytic pathways. With the data presented in this study, PESI-MS can be considered as a potential tool for real-time monitoring of enzymatic reactions because of its simplicity in instrumental configuration, wide applicability under harsh conditions, and flexibility in combination with other techniques. Fil: Yu, Zhan. Shenyang Normal University; China. University Of Yamanashi; Japón Fil: Chen, Lee Chuin. University Of Yamanashi; Japón Fil: Mandal, Mridul Kanti. University Of Yamanashi; Japón Fil: Nonami, Hiroshi. Ehime University; Japón Fil: Erra Balsells, Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Centro de Investigaciones en Hidratos de Carbono. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Centro de Investigaciones en Hidratos de Carbono; Argentina Fil: Hiraoka, Kenzo. University Of Yamanashi; Japón |
description |
Although there are a lot of well established methods for monitoring enzyme-catalyzed reactions, most of them are based on changes in spectroscopic properties during the conversion of substrates to products. However, reactions without optical changes are common, which are inapplicable to these spectroscopic methods. As an alternative technique for enzymologic research, mass spectrometry (MS) is favored due to its specificity, sensitivity, and the ability to obtain stoichiometric information. In this work, probe electrospray ionization (PESI) source coupled with a time of flight mass spectrometer was employed to monitor some typical proteasecatalyzed reactions, including pepsinolysis and trypsinolysis of cytochrome c in real time. Due to the high electrical conductivity of each reaction system, corona discharges are likely to occur, which would decrease intensities of mass spectrometric signals. An ultra-fine sampling probe and an auxiliary vapor spray were adopted to prevent corona discharges. Experimental results from peptic and tryptic digestions of cytochrome c showed different and characteristic catalytic pathways. With the data presented in this study, PESI-MS can be considered as a potential tool for real-time monitoring of enzymatic reactions because of its simplicity in instrumental configuration, wide applicability under harsh conditions, and flexibility in combination with other techniques. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-04 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
format |
article |
status_str |
publishedVersion |
dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/68168 Yu, Zhan; Chen, Lee Chuin; Mandal, Mridul Kanti; Nonami, Hiroshi; Erra Balsells, Rosa; et al.; Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time; Springer; Journal of The American Society for Mass Spectrometry; 23; 4; 4-2012; 728-735 1044-0305 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/68168 |
identifier_str_mv |
Yu, Zhan; Chen, Lee Chuin; Mandal, Mridul Kanti; Nonami, Hiroshi; Erra Balsells, Rosa; et al.; Online electrospray ionization mass spectrometric monitoring of protease-catalyzed reactions in real time; Springer; Journal of The American Society for Mass Spectrometry; 23; 4; 4-2012; 728-735 1044-0305 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/doi/10.1007/s13361-011-0323-5 info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007%2Fs13361-011-0323-5 |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
eu_rights_str_mv |
openAccess |
rights_invalid_str_mv |
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
dc.format.none.fl_str_mv |
application/pdf application/pdf |
dc.publisher.none.fl_str_mv |
Springer |
publisher.none.fl_str_mv |
Springer |
dc.source.none.fl_str_mv |
reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
reponame_str |
CONICET Digital (CONICET) |
collection |
CONICET Digital (CONICET) |
instname_str |
Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.name.fl_str_mv |
CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
repository.mail.fl_str_mv |
dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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1844613800876048384 |
score |
13.070432 |