Production, analysis and bioactivity of recombinant vasoactive intestinal peptide analogs
| Autor: | Yves Cenatiempo, J. Raingeaud, Jean-Marc Muller, Raymond Julien, V. Lelievre, F. Lavergne |
|---|---|
| Rok vydání: | 1996 |
| Předmět: |
Recombinant Fusion Proteins
Molecular Sequence Data Vasoactive intestinal peptide Hydroxylamine In Vitro Techniques Biology Hydroxylamines Biochemistry Cyclase law.invention law Cyclic AMP Animals Humans Amino Acid Sequence Glutathione Transferase chemistry.chemical_classification Base Sequence C-terminus Biological activity General Medicine Fusion protein Molecular biology Amino acid Trachea Oligodeoxyribonucleotides chemistry Recombinant DNA Receptors Vasoactive Intestinal Peptide Rabbits HT29 Cells Linker hormones hormone substitutes and hormone antagonists Vasoactive Intestinal Peptide |
| Zdroj: | Biochimie. 78:14-25 |
| ISSN: | 0300-9084 |
| DOI: | 10.1016/0300-9084(96)81324-1 |
| Popis: | Recombinant vasoactive intestinal polypeptide (VIP) analogs were expressed in Escherichia coli as a fusion protein containing tandemly repeated multiple copies of a synthetic VIP gene joined to glutathione S-transferase. The encoded protein contains VIP units separated by a linker peptide, potentially excisable by a double cleavage with endoprotease factor Xa and hydroxylamine. Expression of different polyVIP genes, from 1 to 32 units, was detected and the production of a 16 VIP polymer was performed. MonoVIP analogs appended by 5 or 10 amino acids at their C terminus were released by factor Xa from this polymerized product. They were then submitted to hydroxylamine cleavage to remove the linker sequence to finally obtain a recombinant VIP analog devoid of any amino acid extension. The biological activity of the recombinant polyVIP and VIP analogs was tested. Although less efficient than the natural neuropeptide, some of these components bound to VIP receptor, activated adenylate cyclase in human colonic adenocarcinoma cells and displayed a relaxation activity on guinea pig tracheal rings. |
| Databáze: | OpenAIRE |
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