Journal
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Volume 27, Issue 9, Pages 1435-1442Publisher
SPRINGER
DOI: 10.1007/s13361-016-1419-8
Keywords
Photodissociation; UVPD; IRMPD; Ubiquitin; Top-down proteomics
Funding
- European Research Council under the European Union's Seventh Framework Program (FP7) [320659]
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Herein we report the successful implementation of the consecutive and simultaneous photodissociation with high (213 nm) and low (10.6 mu m) 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-alpha 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.
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