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
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/68168
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling 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 Técnicas2025-10-22T11:30:37Zoai: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:34982025-10-22 11:30:38.147CONICET 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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score 13.229304

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