| Autor: |
Serra, Esteban C., Krapp, Adriana R., Ottado, Jorgelina, Feldman, Mario F., Ceccarelli, Eduardo A., Carrillo, Néstor |
| Zdroj: |
Journal of Biological Chemistry; August 1995, Vol. 270 Issue: 34 p19930-19935, 6p |
| Abstrakt: |
The precursor of the chloroplast flavoprotein ferredoxin-NADP+reductase from pea was expressed in Escherichia colias a carboxyl-terminal fusion to glutathione S-transferase. The fused protein was soluble, and the precursor could be purified in a few steps involving affinity chromatography on glutathione-agarose, cleavage of the transferase portion by protease Xa, and ion exchange chromatography on DEAE-cellulose. The purified prereductase contained bound FAD but displayed marginally low levels of activity. Removal of the transit peptide by limited proteolysis rendered a functional protease-resistant core exhibiting enzymatic activity. The FAD-containing precursor expressed in E. coliwas readily transported into isolated pea chloroplasts and was processed to the mature size, both inside the plastid and by incubation with stromal extracts in a plastid-free reaction. Import was dependent on the presence of ATP and was stimulated severalfold by the addition of plant leaf extracts. |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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