Determining sequences and post-translational modifications of novel conotoxins inConus victoriae using cDNA sequencing and mass spectrometry
| Autor: | Jonathan V. Sweedler, Ken R. Gayler, Jennifer A. Jakubowski, Bruce G. Livett, Wayne P. Kelley, David A. Keays, Jon-Paul Bingham, David W. Sandall |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
DNA
Complementary Protein mass spectrometry Stereochemistry Electrospray ionization Molecular Sequence Data Mollusk Venoms Peptide Mass spectrometry Mass Spectrometry Animals Amino Acid Sequence Conotoxin Conus victoriae Cloning Molecular Peptide sequence Spectroscopy chemistry.chemical_classification Chromatography Base Sequence Sequence Homology Amino Acid biology Chemistry Sequence Analysis DNA biology.organism_classification Matrix-assisted laser desorption/ionization Conotoxins Protein Processing Post-Translational |
| Zdroj: | Journal of Mass Spectrometry. 39:548-557 |
| ISSN: | 1096-9888 1076-5174 |
| Popis: | A combination of cDNA cloning and detailed mass spectrometric analyses was employed to identify novel conotoxins from Conus victoriae. Eleven conotoxin sequences were determined using molecular methods: one belonging to the A superfamily (Vc1.1), six belonging to the O superfamily (Vc6.1-Vc6.6) and four members of the T superfamily (Vc5.1-Vc5.4). In order to verify the sequences and identify the post-translational modifications (excluding the disulfide connectivity) of three Conus victoriae conotoxins, vc1a, vc5a and vc6a, deduced from sequences Vc1.1, Vc5.1, and Vc6.1, respectively, liquid chromatography/electrospray ionization ion trap mass spectrometry, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and nanospray ionization ion trap mass spectrometry with collisionally induced dissociation were performed on reduced and alkylated venom fractions. We report that vc1a, the native form of alpha-conotoxin Vc1.1 (an unmodified 16 amino acid residue peptide that has notable pain-relieving capabilities), includes a hydroxyproline and a gamma-carboxyglutamate residue. Conotoxin vc5a is a 10-residue peptide with two disulfide bonds and a hydroxyproline and vc6a is a 25 amino acid peptide with three disulfide bonds. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text