RUO Principal

Repositorio Institucional de la Universidad de Oviedo

Ver ítem 
  •   RUO Principal
  • Producción Bibliográfica de UniOvi: RECOPILA
  • Artículos
  • Ver ítem
  •   RUO Principal
  • Producción Bibliográfica de UniOvi: RECOPILA
  • Artículos
  • Ver ítem
    • español
    • English
JavaScript is disabled for your browser. Some features of this site may not work without it.

Listar

Todo RUOComunidades y ColeccionesPor fecha de publicaciónAutoresTítulosMateriasxmlui.ArtifactBrowser.Navigation.browse_issnPerfil de autorEsta colecciónPor fecha de publicaciónAutoresTítulosMateriasxmlui.ArtifactBrowser.Navigation.browse_issn

Mi cuenta

AccederRegistro

Estadísticas

Ver Estadísticas de uso

AÑADIDO RECIENTEMENTE

Novedades
Repositorio
Cómo publicar
Recursos
FAQs

Protein labelling with mercury tags: fundamental studies on ovalbumin derivatised with p-hydroxymercuribenzoic acid (pHMB)

Autor(es) y otros:
Kutscher, Daniel JurgenAutoridad Uniovi; Castillo Busto, María Estela delAutoridad Uniovi; Zinn, Nico; Sanz Medel, AlfredoAutoridad Uniovi; Bettmer, JörgAutoridad Uniovi
Fecha de publicación:
2008
Versión del editor:
http://dx.doi.org/10.1039/b806118a
Citación:
Journal of Analytical Atomic Spectrometry, 23, p. 1359-1364 (2008); doi:10.1039/b806118a
Descripción física:
p. 1359-1364
Resumen:

Protein labelling in combination with mass spectrometry is appointed as a modern approach for quantifying biopolymers, especially proteins. With respect to elemental mass spectrometry, specifically inductively coupled plasma-mass spectrometry (ICP-MS), protein labelling approaches are still scarce, although they offer many advantages, e.g. in terms of detection sensitivity. In this fundamental work, we present results on the labelling of ovalbumin with p-hydroxymercuribenzoic acid (pHMB). After optimising the derivatisation procedure, the characterisation of the labelled species is necessary, and thus, the use of molecular MS techniques like MALDI-, and ESI-MS is required. Finally, the detection capabilities of ICP-MS are evaluated on the labelled species. Important factors to consider are the reaction yield, the selectivity, and the stoichiometry of the bioconjugate. For instance, the stoichiometry of the bioconjugate is determined by comparative measurements using MALDI-, and ESI-MS. It can be demonstrated that the label/protein ratio is determined to be [similar]3 : 1 by MALDI-MS, which is lower than the number of expected binding sites (ovalbumin has four free sulfhydryl groups from cysteines). In contrast to these findings, the use of ESI-Q-ToF-MS with its superior mass resolution indicates a stoichiometry of 4 : 1. However, the overall strategy given here on the example of ovalbumin labelling with pHMB might be a promising approach for protein quantification as it provides a significant improvement in terms of detection limits (1 fmol for ovalbumin) in comparison to the use of sulfur as naturally occurring elemental tag.

Protein labelling in combination with mass spectrometry is appointed as a modern approach for quantifying biopolymers, especially proteins. With respect to elemental mass spectrometry, specifically inductively coupled plasma-mass spectrometry (ICP-MS), protein labelling approaches are still scarce, although they offer many advantages, e.g. in terms of detection sensitivity. In this fundamental work, we present results on the labelling of ovalbumin with p-hydroxymercuribenzoic acid (pHMB). After optimising the derivatisation procedure, the characterisation of the labelled species is necessary, and thus, the use of molecular MS techniques like MALDI-, and ESI-MS is required. Finally, the detection capabilities of ICP-MS are evaluated on the labelled species. Important factors to consider are the reaction yield, the selectivity, and the stoichiometry of the bioconjugate. For instance, the stoichiometry of the bioconjugate is determined by comparative measurements using MALDI-, and ESI-MS. It can be demonstrated that the label/protein ratio is determined to be [similar]3 : 1 by MALDI-MS, which is lower than the number of expected binding sites (ovalbumin has four free sulfhydryl groups from cysteines). In contrast to these findings, the use of ESI-Q-ToF-MS with its superior mass resolution indicates a stoichiometry of 4 : 1. However, the overall strategy given here on the example of ovalbumin labelling with pHMB might be a promising approach for protein quantification as it provides a significant improvement in terms of detection limits (1 fmol for ovalbumin) in comparison to the use of sulfur as naturally occurring elemental tag.

URI:
http://hdl.handle.net/10651/8927
ISSN:
0267-9477
Identificador local:

319

DOI:
10.1039/b806118a
Colecciones
  • Artículos [37532]
Ficheros en el ítem
Métricas
Compartir
Exportar a Mendeley
Estadísticas de uso
Estadísticas de uso
Metadatos
Mostrar el registro completo del ítem
Página principal Uniovi

Biblioteca

Contacto

Facebook Universidad de OviedoTwitter Universidad de Oviedo
El contenido del Repositorio, a menos que se indique lo contrario, está protegido con una licencia Creative Commons: Attribution-NonCommercial-NoDerivatives 4.0 Internacional
Creative Commons Image