An EThcD-Based Method for Discrimination of Leucine and Isoleucine Residues in Tryptic Peptides
| Autor: | Sergey V. Kovalyov, Tatiana Yu. Samgina, S. S. Zhokhov, Albert T. Lebedev |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Electrospray ionization Serum Albumin Human Peptide Mass spectrometry Orbitrap 01 natural sciences Amphibian Proteins Mass Spectrometry law.invention 03 medical and health sciences Leucine Sequence Analysis Protein Structural Biology law medicine Animals Humans Amino Acid Sequence Isoleucine Rana ridibunda Spectroscopy chemistry.chemical_classification Chromatography Edman degradation Chemistry 010401 analytical chemistry Human serum albumin Peptide Fragments 0104 chemical sciences 030104 developmental biology Biochemistry Antimicrobial Cationic Peptides medicine.drug |
| Zdroj: | Journal of the American Society for Mass Spectrometry. 28:1600-1611 |
| ISSN: | 1044-0305 |
| DOI: | 10.1007/s13361-017-1674-3 |
| Popis: | An EThcD-based approach for the reliable discrimination of isomeric leucine and isoleucine residues in peptide de novo sequencing procedure has been proposed. A multistage fragmentation of peptide ions was performed with Orbitrap Elite mass spectrometer in electrospray ionization mode. At the first stage, z-ions were produced by ETD or ETcaD fragmentation of doubly or triply charged peptide precursor ions. These primary ions were further fragmented by HCD with broad-band ion isolation, and the resulting w-ions showed different mass for leucine and isoleucine residues. The procedure did not require manual isolation of specific z-ions prior to HCD stage. Forty-three tryptic peptides (3 to 27 residues) obtained by trypsinolysis of human serum albumin (HSA) and gp188 protein were analyzed. To demonstrate a proper solution for radical site migration problem, three non-tryptic peptides were also analyzed. A total of 93 leucine and isoleucine residues were considered and 83 of them were correctly identified. The developed approach can be a reasonable substitution for additional Edman degradation procedure, which is still used in peptide sequencing for leucine and isoleucine discrimination. Graphical Abstract ᅟ. |
| Databáze: | OpenAIRE |
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