Cargando…

Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization

Herein we report the successful implementation of the consecutive and simultaneous photodissociation with high (213 nm) and low (10.6 μm) energy photons (HiLoPD, high-low photodissociation) on ubiquitin in a quadrupole-Orbitrap mass spectrometer. Absorption of high-energy UV photon is dispersed over...

Descripción completa

Detalles Bibliográficos
Autores principales: Halim, Mohammad A., Girod, Marion, MacAleese, Luke, Lemoine, Jérôme, Antoine, Rodolphe, Dugourd, Philippe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5031736/
https://www.ncbi.nlm.nih.gov/pubmed/27287047
http://dx.doi.org/10.1007/s13361-016-1419-8
_version_ 1782454847348408320
author Halim, Mohammad A.
Girod, Marion
MacAleese, Luke
Lemoine, Jérôme
Antoine, Rodolphe
Dugourd, Philippe
author_facet Halim, Mohammad A.
Girod, Marion
MacAleese, Luke
Lemoine, Jérôme
Antoine, Rodolphe
Dugourd, Philippe
author_sort Halim, Mohammad A.
collection PubMed
description Herein we report the successful implementation of the consecutive and simultaneous photodissociation with high (213 nm) and low (10.6 μm) energy photons (HiLoPD, high-low photodissociation) on ubiquitin in a quadrupole-Orbitrap mass spectrometer. Absorption of high-energy UV photon is dispersed over the whole protein and stimulates extensive C–C(α) backbone fragmentation, whereas low-energy IR photon gradually increases the internal energy and thus preferentially dissociates the most labile amide (C–N) bonds. We noticed that simultaneous irradiation of UV and IR lasers on intact ubiquitin in a single MS/MS experiment provides a rich and well-balanced fragmentation array of a/x, b/y, and z ions. Moreover, secondary fragmentation from a/x and z ions leads to the formation of satellite side-chain ions (d, v, and w) and can help to distinguish isomeric residues in a protein. Implementation of high-low photodissociation in a high-resolution mass spectrometer may offer considerable benefits to promote a comprehensive portrait of protein characterization. [Figure: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s13361-016-1419-8) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-5031736
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Springer US
record_format MEDLINE/PubMed
spelling pubmed-50317362016-10-09 Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization Halim, Mohammad A. Girod, Marion MacAleese, Luke Lemoine, Jérôme Antoine, Rodolphe Dugourd, Philippe J Am Soc Mass Spectrom Research Article Herein we report the successful implementation of the consecutive and simultaneous photodissociation with high (213 nm) and low (10.6 μm) energy photons (HiLoPD, high-low photodissociation) on ubiquitin in a quadrupole-Orbitrap mass spectrometer. Absorption of high-energy UV photon is dispersed over the whole protein and stimulates extensive C–C(α) backbone fragmentation, whereas low-energy IR photon gradually increases the internal energy and thus preferentially dissociates the most labile amide (C–N) bonds. We noticed that simultaneous irradiation of UV and IR lasers on intact ubiquitin in a single MS/MS experiment provides a rich and well-balanced fragmentation array of a/x, b/y, and z ions. Moreover, secondary fragmentation from a/x and z ions leads to the formation of satellite side-chain ions (d, v, and w) and can help to distinguish isomeric residues in a protein. Implementation of high-low photodissociation in a high-resolution mass spectrometer may offer considerable benefits to promote a comprehensive portrait of protein characterization. [Figure: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s13361-016-1419-8) contains supplementary material, which is available to authorized users. Springer US 2016-06-10 2016 /pmc/articles/PMC5031736/ /pubmed/27287047 http://dx.doi.org/10.1007/s13361-016-1419-8 Text en © American Society for Mass Spectrometry 2016
spellingShingle Research Article
Halim, Mohammad A.
Girod, Marion
MacAleese, Luke
Lemoine, Jérôme
Antoine, Rodolphe
Dugourd, Philippe
Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title_full Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title_fullStr Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title_full_unstemmed Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title_short Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title_sort combined infrared multiphoton dissociation with ultraviolet photodissociation for ubiquitin characterization
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5031736/
https://www.ncbi.nlm.nih.gov/pubmed/27287047
http://dx.doi.org/10.1007/s13361-016-1419-8
work_keys_str_mv AT halimmohammada combinedinfraredmultiphotondissociationwithultravioletphotodissociationforubiquitincharacterization
AT girodmarion combinedinfraredmultiphotondissociationwithultravioletphotodissociationforubiquitincharacterization
AT macaleeseluke combinedinfraredmultiphotondissociationwithultravioletphotodissociationforubiquitincharacterization
AT lemoinejerome combinedinfraredmultiphotondissociationwithultravioletphotodissociationforubiquitincharacterization
AT antoinerodolphe combinedinfraredmultiphotondissociationwithultravioletphotodissociationforubiquitincharacterization
AT dugourdphilippe combinedinfraredmultiphotondissociationwithultravioletphotodissociationforubiquitincharacterization