Direct expression in Escherichia coli and characterization of bovine adrenodoxins with modified amino‐terminal regions

FEBS Letters

Ariyasu, Yasuyuki²  Horiuchi, Tadao²  Ando, Fumiko²  Hara, Takayuki¹  Tokunaga, Naomi²  Sagara, Yasuhiro² 

[1] Department of Food and Nutrition, Nakamura Gakuen College, Jonan-ku, Fukuoka 814-01, Japan;Department of Microbiology, Faculty of Pharmaceutical Sciences, Kyushu University, Higashi-ku, Fukuoka 812, Japan

关键词: Adrenodoxin;    Adrenodoxin Reductase;    cDNA;    Cytochrome c;    Cytochrome P-450SCC;    Expression of protein;    Ad;    adrenodoxin;    AdR;    NADPH-adrenodoxin reductase;    M-Ad;    mature form of bovine adrenocortical adrenodoxin;    P-450SCC;    cytochrome P-450 catalyzing;    cholesterol side chain cleavage reaction;    P-45011β;    cytochrome P-150 catalyzing steroid 11β-hydroxylation;    ITPG;    isopropyl-β-d-thiogalactopyranoside;    SDS-PAGE;    sodium dodecyl sulfate/polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(92)80847-A

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Four forms of bovine adrenodoxin with modified amino-termini obtained by direct expression of cDNAs in Escherichia coli are Ad(Met¹), Ad(Met⁻¹), Ad(Met⁻¹²), and Ad(Met⁶). The shoulder numbers represent this site of translation initiator Met at the amino-termini. The adrenodoxins, except for Ad(Met⁻¹), were purified from the cell lysate and the ratios of A ₄₁₄-to-A ₂₇₆ of the purified proteins were over 0.92. NADPH-cytochrome c reductase activities of the three forms of adrenodoxin in the presence of adrenodoxin reductase were the same as that of purified bovine adrenocortical adrenodoxin. However, as cytochrome P-450_SCC reduction catalyzed by Ad(Met⁰) was about 60% or that by Ad(Met¹), the contribution of the amino-terminal region for the electron transfer or binding to cytochrome P-450_SCC would need to be considered.

【 授权许可】

Unknown