Expression and analysis of recombinant Amb a V and Amb t V allergens. Comparison with native proteins by immunological assays and NMR spectroscopy
| Autor: | T Rafnar, Shau-Ku Huang, Balaram Ghosh, L. Mueller, David G. Marsh, M. P. Perry, W. J. Metzler |
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| Rok vydání: | 1992 |
| Předmět: |
Ragweed
Stereochemistry Biological activity Radioimmunoassay Cell Biology Nuclear magnetic resonance spectroscopy Biology medicine.disease_cause biology.organism_classification Biochemistry Molecular biology law.invention Allergen Immune system law Recombinant DNA medicine Molecular Biology Escherichia coli |
| Zdroj: | Journal of Biological Chemistry. 267:21119-21123 |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/s0021-9258(19)36805-x |
| Popis: | The Amb V allergens are small, highly disulfide-bonded ragweed pollen allergens that serve as useful models for understanding the molecular basis of the human immune response. We have produced recombinant Amb a V and Amb t V (from short and giant ragweed pollens, respectively) in Escherichia coli and have compared their structural and functional characteristics to those of the native proteins. Recombinant Amb t V was indistinguishable from native Amb t V as determined by NMR spectroscopy and antibody-binding studies. Whereas inhibition analysis showed that recombinant Amb a V possessed only approximately 50% of the antibody-binding activity of native Amb a V, the two proteins were similarly effective in stimulating Amb a V-specific T-cells. Our results demonstrate that even highly homologous proteins exhibit different abilities to fold into their native three-dimensional conformations and establish the potential and limits of expressing the recombinant Amb V allergens intracellularly in E. coli. |
| Databáze: | OpenAIRE |
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