Production of the R2 subunit of ribonucleotide reductase from herpes simplex virus with prokaryotic and eukaryotic expression systems: higher activity of R2 produced by eukaryotic cells related to higher iron-binding capacity
| Autor: | Bernard Massie, F Dumas, Marc Fontecave, Y. Langelier, P. Gaudreau, G Matton, Mohamed G. Atta, Nathalie Lamarche |
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| Rok vydání: | 1996 |
| Předmět: |
Herpesvirus 2
Human Iron Protein subunit Dimer medicine.disease_cause Biochemistry Cofactor Cell Line law.invention chemistry.chemical_compound law Cricetinae Ribonucleotide Reductases medicine pharmaceutical Animals Prokaryotic expression Cloning Molecular Molecular Biology biology Cell Biology Molecular biology Recombinant Proteins Ribonucleotide reductase Herpes simplex virus chemistry biology.protein Recombinant DNA Tyrosine Electrophoresis Polyacrylamide Gel Iron binding capacity Protein Binding Research Article |
| Zdroj: | Biochemical Journal. 320:129-135 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj3200129 |
| Popis: | The R2 subunit of ribonucleotide reductase from herpes simplex virus type 2 was overproduced with prokaryotic and eukaryotic expression systems. The recombinant R2 purified by a two-step procedure exhibited a 3-fold higher activity when produced in eukaryotic cells. Precise quantification of the R2 concentration at each step of the purification indicated that the activity was not altered during the purification procedure. Moreover, we have observed that the level of R2 expression, in eukaryotic cells as well as in prokaryotic cells, did not influence R2 activity. Extensive characterization of the recombinant R2 purified from eukaryotic and prokaryotic expression systems has shown that both types of pure R2 preparations were similar in their 76 kDa dimer contents (more than 95%) and in their ability to bind the R1 subunit. However, we have found that the higher activity of R2 produced in eukaryotic cells is more probably related to a higher capability of binding the iron cofactor as well as a 3-fold greater ability to generate the tyrosyl free radical. |
| Databáze: | OpenAIRE |
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