Establishing Signature Fragments for Identification and Sequencing of Dityrosine Cross-Linked Peptides Using Ultraviolet Photodissociation Mass Spectrometry
| Autor: | Eugene A. Kapp, Craig A. Hutton, Soumya Mukherjee, Blaine R. Roberts, Mengxuan Fang, Romain Huguet, Varsha J. Thombare, W. Mei Kok, Gavin E. Reid |
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| Rok vydání: | 2019 |
| Předmět: |
chemistry.chemical_classification
Amyloid beta-Peptides Ultraviolet Rays Stereochemistry Sequence analysis 010401 analytical chemistry Photodissociation Peptide Photochemical Processes 010402 general chemistry Mass spectrometry Tandem mass spectrometry Cleavage (embryo) 01 natural sciences Peptide Fragments 0104 chemical sciences Analytical Chemistry Residue (chemistry) chemistry Fragmentation (mass spectrometry) Sequence Analysis Protein Tandem Mass Spectrometry Tyrosine Peptides Protein Processing Post-Translational |
| Zdroj: | Analytical Chemistry. 91:12129-12133 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.9b02986 |
| Popis: | Dityrosine cross-linking of Aβ peptides and α-synuclein is increasingly becoming recognized as a biomarker of neuropathological diseases. However, there remains a need for the development of analytical methods that enable the specific and selective identification of dityrosine cross-linked proteins and peptides in complex biological samples. Here, we report that the gas-phase fragmentation of protonated dityrosine cross-linked peptides under ultraviolet photodissociation (UVPD) tandem mass spectrometry (MS/MS) conditions results in the cleavage across Cα and Cβ atoms of the dityrosine residue. This Cα-Cβ cleavage in UVPD-MS/MS results in the formation of diagnostic pairs of product ions, providing information on the two individual peptides involved in the cross-linking, resolving the intrinsic "n2 problem" plaguing the identification of this post-translational modification (PTM) by tandem mass spectrometry. Sequencing of a heterodimeric dityrosine cross-linked peptide was demonstrated using hybrid UVPD-MS/MS and CID-MS3 on a diagnostic pair of product ions. In combination with dedicated MS-cleavable MSn software, UVPD-MSn therefore provides an avenue to selectively discover and describe dityrosine cross-linked peptides. Additionally, observation of dityrosine-specific "reporter ions" at m/z 240.1019 and m/z 223.0752 in UVPD-MS/MS will be useful for the validation of the dityrosine cross-linked peptides. |
| Databáze: | OpenAIRE |
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