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• W2753113281 abstract "Rationale Tandem mass spectrometry (MS/MS) is the pivotal tool for protein structural characterization and quantification. Identification relies on the fragmentation step of tryptic peptides in bottom‐up strategy. Specificity of fragmentation can be obtained using laser‐induced dissociation (LID) in the visible range, after tagging of the targeted peptides with an adequate chromophore. Backbone fragmentation is required to obtain specific fragments and confident identification. We present herein a study of fragmentation patterns of chromophore‐tagged peptides in LID, showing the potential of LID methodology to provide the maximum number of fragments for further identification and quantification. Methods A total of 401 cysteine‐containing tryptic peptides originating from the human proteome were derivatizated on the thiol group of cysteine with a Dabcyl maleimide chromophore, which has a high photo‐absorption cross section at 473 nm. The derivatized peptides were then analyzed by LID at 473 nm on a Q Exactive instrument. Results LID spectra present a characteristic fragment at m/z 252.112 for all precursors. This product ion arises from the internal dissociation of the Dabcyl chromophore. Several peptide‐backbone fragment ions are also detected. Results show the quasi absence of fragmentation at the cysteine site. This indicates that part of the energy must be redistributed across the entire system despite excitation initially localized at the chromophore. Indeed, the fragmentation mainly occurs at 3 to 5 amino acids from the derivatized cysteine residue. Conclusions LID of derivatized cysteine‐containing peptides displays the initial fragmentation of the chromophore. As energy is redistributed all along the peptide sequence, fragmentation of the peptide backbone is also observed. Thus, LID of chromophore‐tagged peptides produces adequate fragment ions, allowing both good sequence coverage for a greater confidence of identification, and a large choice of transitions for specific quantification." @default.
• W2753113281 created "2017-09-15" @default.
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• W2753113281 date "2017-10-29" @default.
• W2753113281 modified "2023-10-16" @default.
• W2753113281 title "Fragmentation patterns of chromophore-tagged peptides in visible laser induced dissociation" @default.
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• W2753113281 doi "https://doi.org/10.1002/rcm.7984" @default.
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