Direct expression in Escherichia coli and characterization of bovine adrenodoxins with modified amino-terminal regions.
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 the 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 A414-to-A276 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-450SCC reduction catalyzed by Ad(Met6) was about 60% of that by Ad(Met1), the contribution of the amino-terminal region for the electron transfer or binding to cytochrome P-450SCC would need to be considered.
Sagara
Y
Y
Department of Microbiology, Faculty of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
Hara
T
T
Ariyasu
Y
Y
Ando
F
F
Tokunaga
N
N
Horiuchi
T
T
D000325:Adrenodoxin / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D002417:Cattle; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry* / Q000302:isolation & purification; D013329:Structure-Activity Relationship
D011994:Recombinant Proteins; D000325:Adrenodoxin