Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization

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...

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Main Authors: Halim, Mohammad A., Girod, Marion, MacAleese, Luke, Lemoine, Jérôme, Antoine, Rodolphe, Dugourd, Philippe
Format: Online
Language:English
Published: Springer US 2016
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5031736/
id pubmed-5031736
recordtype oai_dc
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 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–Cα 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. 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
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Halim, Mohammad A.
Girod, Marion
MacAleese, Luke
Lemoine, Jérôme
Antoine, Rodolphe
Dugourd, Philippe
spellingShingle Halim, Mohammad A.
Girod, Marion
MacAleese, Luke
Lemoine, Jérôme
Antoine, Rodolphe
Dugourd, Philippe
Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
author_facet Halim, Mohammad A.
Girod, Marion
MacAleese, Luke
Lemoine, Jérôme
Antoine, Rodolphe
Dugourd, Philippe
author_sort Halim, Mohammad A.
title Combined Infrared Multiphoton Dissociation with Ultraviolet Photodissociation for Ubiquitin Characterization
title_short 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_sort combined infrared multiphoton dissociation with ultraviolet photodissociation for ubiquitin characterization
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–Cα 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.
publisher Springer US
publishDate 2016
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5031736/
_version_ 1613657362229362688

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