Photocross-Linked Peptide-Protein Complexes Analysis: A Comparative Study of CID and ETD Fragmentation Modes
In: Journal of The American Society for Mass Spectrometry Journal of The American Society for Mass Spectrometry, Springer Verlag (Germany), 2015, 26 (6), pp.1014-1026. ⟨10.1007/s13361-015-1095-0⟩ Journal of The American Society for Mass Spectrometry, 2015, 26 (6), pp.1014-1026. ⟨10.1007/s13361-015-1095-0⟩; (2015-04-04)
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Zugriff:
International audience; Protein–protein interactions are among the keys to organizing cellular processes in space and time. One of the only direct ways to identify such interactions in their cellular environment is to covalently bond the interacting partners to fix the interaction. Photocross-linking in living cells is thus a very promising technique. The feasibility of in cellulo photocross-linking reactions has been shown and mass spectrometry is a tool of choice to analyze photocross-linked proteins. However, the interpretation of the MS and MS/MS spectra of photocross-linked peptides remains one of the most important bottlenecks of the method and still limits its potential for large-scale applications (interactomics). Fundamental studies are still necessary to understand and characterize the fragmentation behavior of photocross-linked peptides. Here, we report the successful identification of the interaction sites in a well-characterized model of in vitro interaction between a protein and a peptide. We describe in detail the fragmentation pattern of these photocross-linked species in order to identify trends that could be generalized. In particular, we compare CID and ETD fragmentation modes (and HCD in a lesser extent), demonstrating the complementarity of both methods and the advantage of ETD for the analysis of photocross-linked species. The information should help further development of dedicated software to properly score MS/MS spectra of photocross-linked species.
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Photocross-Linked Peptide-Protein Complexes Analysis: A Comparative Study of CID and ETD Fragmentation Modes
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Autor/in / Beteiligte Person: | Clavier, Séverine ; Sachon, Emmanuelle ; Bolbach, Gérard ; Spectrométrie de Masse et Protéomique [IBPS] (IBPS-SPM) ; Institut de Biologie Paris Seine (IBPS) ; Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) ; Analyse, Interactions Moléculaires et Cellulaires (LBM-E2) ; Laboratoire des biomolécules (LBM UMR 7203) ; Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris ; École normale supérieure - Paris (ENS Paris) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS Paris) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Chimie - ENS Paris ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC) ; Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris ; École normale supérieure - Paris (ENS-PSL) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL) ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Département de Chimie - ENS Paris ; Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS) |
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Quelle: | Journal of The American Society for Mass Spectrometry Journal of The American Society for Mass Spectrometry, Springer Verlag (Germany), 2015, 26 (6), pp.1014-1026. ⟨10.1007/s13361-015-1095-0⟩ Journal of The American Society for Mass Spectrometry, 2015, 26 (6), pp.1014-1026. ⟨10.1007/s13361-015-1095-0⟩; (2015-04-04) |
Veröffentlichung: | HAL CCSD, 2015 |
Medientyp: | unknown |
ISSN: | 1044-0305 (print) ; 1879-1123 (print) |
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