IRMPD and ECD fragmentation of intermolecular cross-linked peptides
| Autor: | Marcos N. Eberlin, Luiz Fernando Arruda Santos, Fabio C. Gozzo |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
chemistry.chemical_classification
Electron-capture dissociation Chemistry Stereochemistry Molecular Sequence Data Analytical chemistry Peptide Mass spectrometry Tandem mass spectrometry Dissociation (chemistry) Mass Spectrometry Fragmentation (mass spectrometry) Sequence Analysis Protein Spectroscopy Fourier Transform Infrared Protein folding Infrared multiphoton dissociation Amino Acid Sequence Peptides Spectroscopy |
| Zdroj: | Journal of mass spectrometry : JMS. 46(3) |
| ISSN: | 1096-9888 |
| Popis: | Despite the increasing number of studies using mass spectrometry for three dimensional analyses of proteins (MS3D), the identification of cross-linked peptides remains a bottleneck of the method. One of the main reasons for this is the lack of knowledge about the fragmentation of these species. Intermolecular cross-linked peptides are considered the most informative species present in MS3D experiment, since different peptides are connected by a cross-linker, the peptides chain can be either from a single protein, providing information about protein folding, or from two different proteins in a complex, providing information about binding partners, complex topology and interaction sites. These species tend to be large and highly charged in ESI, making comprehensive fragmentation by CID MS/MS problematic. On the other hand, these highly charged peptides are very suitable for dissociation using both infrared multiphoton dissociation (IRMPD) and electron capture dissociation (ECD). Herein, we report the fragmentation study of intermolecular cross-linked peptides using IRMPD and ECD. Using synthetic peptides and different commercial cross-linkers, a series of intermolecular cross-linked peptides were generate, and subsequently fragmented by IRMPD and ECD in a FT-ICR-MS instrument. Due to the high mass accuracy and resolution of the FT-ICR, the fragment ions could be attributed with high confidence. The peptides sequence coverage and fragmentation features obtained from IRMPD and ECD were compared for all charge states. |
| Databáze: | OpenAIRE |
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