Overproduction in Escherichia coli and Characterization of the Precise Mature Form of Rat Adrenodoxin

概要

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The mature form of rat adrenodoxin (Ad) was purified from a heterologous direct expression system in Es-cherichia coli with a yield of 56 mg/l culture. The purified Ad showed a A_<414>/A_<280> ratio of 0.91 and the sequence of 10 amino terminal residues was identical with that of authentic rat Ad. By Time of Flight/Mass spectrometry, the molecular mass of purified Ad was identical to that calculated from the cDNA sequence and the carboxy terminal residue was estimated to be Ser, which was also as expected from the cDNA. These results indicate that the purified recombinant Ad is a precise mature form. In measurements of NADPH-cytochrome c reductase activity reconstituted which bovine adrenodoxin reductase (AdR), the apparent K_m value for rat Ad was 46.9±2.5 nM, indicating a somewhat lower affinity for rat Ad to bovine AdR than for bovine Ad. On the other hand, the spectral K_d value for rat Ad to bovine cytochrome P-450scc was 0.46±0.05 μM, a value which was almost identical with that of the bovine counterpart.
公益社団法人日本薬学会の論文
1998-10-15