A 15-kD Interferon-Induced Protein and Its 17-kD Precursor: Expression inEscherichia coli, Purification, and Characterization
| Autor: | Barbara Larsen, Diana Fahey, Dale C. Blomstrom, Neil Feltham, Ernest Knight, Milton Hillman, Beverly Cordova |
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| Rok vydání: | 1989 |
| Předmět: |
Immunology
Biology Molecular cloning medicine.disease_cause law.invention Bacterial Proteins FLAG-tag law Virology Protein A/G Escherichia coli medicine Humans Amino Acid Sequence Protein Precursors Peptide sequence chemistry.chemical_classification Molecular biology Recombinant Proteins Amino acid Molecular Weight Biochemistry chemistry biology.protein Recombinant DNA Interferons Crystallization Myc-tag |
| Zdroj: | Journal of Interferon Research. 9:493-507 |
| ISSN: | 0197-8357 |
| DOI: | 10.1089/jir.1989.9.493 |
| Popis: | Using recombinant DNA technology, a 15-kD interferon (IFN)-induced protein and its 17-kD precursor have been expressed in Escherichia coli to obtain sufficient quantities of each protein for the investigation of their biological roles. Both the 15-kD and 17-kD proteins have been purified to homogeneity and crystallized. The recombinant 15-kD protein has an identical reversed-phase HPLC elution profile to that of the native 15-kD protein purified from human cells. Furthermore, the recombinant 15-kD and 17-kD proteins have identical amino- and carboxy-terminal amino acid sequences to those predicted from the DNA sequence. The native and recombinant 15-kD proteins give identical tryptic peptide maps, and the recombinant 17-kD protein gives only one additional tryptic peptide. We conclude that the recombinant 17-kD and 15-kD proteins are identical to the 17-kD precursor and the 15-kD stable product synthesized in human cells in response to IFN stimulation. In addition, we have demonstrated that the recombinant 17-kD precursor protein can be converted to the 15-kD protein by cytoplasmic extracts of human cells. |
| Databáze: | OpenAIRE |
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