An unusual cleavage reaction of a peptide observed during dithiotreitol and tris(2-carboxyethyl)phosphine reduction: application to sequencing of HpTx2 spider toxin using nanospray tandem mass spectrometry
| Autor: | Marie-Louise Célérier, Christian Legros, Catherine Guette |
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| Rok vydání: | 2004 |
| Předmět: |
Spectrometry
Mass Electrospray Ionization Sequence analysis Phosphines Molecular Sequence Data Spider Venoms Peptide medicine.disease_cause Tandem mass spectrometry Mass spectrometry Analytical Chemistry chemistry.chemical_compound Sequence Analysis Protein medicine Nanotechnology Amino Acid Sequence Peptide sequence Spectroscopy chemistry.chemical_classification Chromatography Chemistry Toxin Organic Chemistry Spider toxin Dithiothreitol TCEP Peptides Oxidation-Reduction |
| Zdroj: | Rapid communications in mass spectrometry : RCM. 18(12) |
| ISSN: | 0951-4198 |
| Popis: | A recombinant peptidic spider toxin, HpTx2, was investigated directly by nanoelectrospray tandem mass spectrometry (MS/MS). This 30-residue toxin possesses a highly knotted structure with cystines arranged in close proximity. The low-energy collision-induced dissociation MS/MS spectrum of the [M+4H](4+) ion permitted characterization of the C-terminal sequence of HpTx2 up to Cys(26) that is involved in a disulfide bridge. Chemical pre-treatment with DTT or TCEP was then investigated, and it was found that an unexpected cleavage reaction of HpTx2 gave two smaller peptides which were completely sequenced by MS/MS experiments using a Qq-TOF mass spectrometer. This unusual hydrolysis reaction facilitated the determination of the complete sequence of the HpTx2 toxin. |
| Databáze: | OpenAIRE |
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