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标题：Overproduction in Escherichia coli and Characterization of the Precise Mature Form of Rat Adrenodoxin
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作者：Yasuhiro SAGARA ; Hironori ARAMAKi
期刊名称：Biological and Pharmaceutical Bulletin
印刷版ISSN：0918-6158
电子版ISSN：1347-5215
出版年度：1998
卷号：21
期号：10
页码：1106-1109
DOI：10.1248/bpb.21.1106
出版社：The Pharmaceutical Society of Japan
摘要：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 A₄₁₄/A₂₈₀ 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.
关键词：adrenodoxin;adrenodoxin reductase;cDNA;cytochrome P-450scc;Escherichia coli;rat