Characterization and location of post-translational modifications on chromogranin B from bovine adrenal medullary chromaffin granules
| Autor: | Claire Gasnier, Olivier Ruh, Jean-Marc Strub, Marie-Hélène Metz-Boutigue, Dominique Aunis, Karine Lugardon |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Proteomics
Glycosylation Proteome Amino Acids Acidic Molecular Sequence Data Biochemistry chemistry.chemical_compound Western blot Liquid chromatography–mass spectrometry Sequence Analysis Protein medicine Chromogranins Animals Chromaffin Granules Amino Acid Sequence Molecular Biology Gel electrophoresis biology Molecular mass medicine.diagnostic_test Sequence Homology Amino Acid Chromogranin A Molecular biology Protein Structure Tertiary Molecular Weight Isoelectric point medicine.anatomical_structure chemistry Adrenal Medulla Mutation biology.protein Cattle Adrenal medulla Protein Processing Post-Translational |
| Zdroj: | Proteomics. 4(6) |
| ISSN: | 1615-9853 |
| Popis: | Bovine chromoganin B (CGB)/secretogranin I, an acidic protein with a sequence of 626 residues and an isoelectric point of 5.2 is a major member of the chromogranin/secretogranin (CG/Sg) family. The difference between the theoretical molecular mass (76 kDa) and the value estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis results from post-translational modifications (glycosylation, phosphorylation and sulfation) and from the abundance of acidic residues (D 4.6%, and E 16.5%). Although the sequence of CGB is known, the structural analyses of the post-translational modifications have so far not been carried out. In the present study, using a combination of proteomic techniques including two-dimensional gel electrophoresis, Western blot, high-performance liquid chromatography purification, enzymatic digestion, sequencing, carbohydrate analysis, matrix-assisted laser desorption/ionization-time of flight and liquid chromatography mass spectrometry analysis, we have located 18 post-translational modifications on bovine CGB, isolated from adrenal medulla chromaffin granules. Furthermore, we have identified at the molecular level the presence of a mutation M/V on position 577 of natural CGB. All together these data reflect the complex structure of this protein marker of the neuroendocrine system. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text