Novel cysteine tags for the sequencing of non-tryptic disulfide peptides of anurans: ESI-MS study of fragmentation efficiency
| Autor: | Egor Vorontsov, Basem Kanawati, Konstantin A. Artemenko, Ilya E. Nifant'ev, Roman A. Zubarev, Philippe Schmitt-Kopplin, Tatyana Yu. Samgina, Albert T. Lebedev, Vladimir Gorshkov |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Spectrometry Mass Electrospray Ionization Iodoacetic acid Sequence analysis Electrospray ionization Molecular Sequence Data Orbitrap Mass spectrometry Amphibian Proteins law.invention Maleimides chemistry.chemical_compound Structural Biology law Sequence Analysis Protein Tandem Mass Spectrometry Animals Amino Acid Sequence Cysteine Disulfides Maleimide Peptide sequence Spectroscopy Chromatography High Pressure Liquid Rana ridibunda Skin Chromatography Iodoacetic Acid chemistry Molecular Probes Female Molecular probe Antimicrobial Cationic Peptides |
| Zdroj: | Journal of the American Society for Mass Spectrometry. 22(12) |
| ISSN: | 1879-1123 |
| Popis: | Mass spectrometry faces considerable difficulties in de novo sequencing of long non-tryptic peptides with S–S bonds. Long disulfide-containing peptides brevinins 1E and 2Ec from frog Rana ridibunda were reduced and alkylated with nine novel and three known derivatizing agents. Eight of the novel reagents are maleimide derivatives. Modified samples were subjected to MS/MS studies on FT-ICR and Orbitrap mass spectrometers using CAD/HCD or ECD/ETD techniques. Procedures, fragmentation patterns, and sequence coverage for two peptides modified with 12 tags are described. ECD/ETD and CAD fragmentation revealed complementary sequence information. Higher-energy collisionally activated dissociation (HCD) sufficiently enhanced y-ions formation for brevinin 1E, but not for brevinin 2Ec. Some novel tags [N-benzylmaleimide, N-(2,6-dimethylphenyl)maleimide] along with known N-phenylmaleimide and iodoacetic acid showed high total sequence coverage taking into account combined ETD and HCD fragmentation. Moreover, modification of long (34 residues) brevinin 2Ec with N-benzylmaleimide or N-(2,6-dimethylphenyl)maleimide yielded high sequence coverage and full C-terminal sequence determination with ECD alone. |
| Databáze: | OpenAIRE |
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