Chaperone-assisted expression of authentic bovine adrenodoxin reductase in Escherichia coli.
Adrenodoxin reductase is an essential component of the mitochondrial monooxygenase systems that are involved in the synthesis of steroid hormones and related compounds. After removing by mutagenesis a secondary ribosome binding site and an mRNA loop formed between the gene and the vector, large amounts of the enzyme could be produced in Escherichia coli by coexpression with the HSP60-chaperone system. The purified protein was homogeneous enough for reproducible crystallization. The crystals diffracted X-rays isotropically beyond 1.7 A resolution permitting a structure analysis.
Vonrhein
C
C
Institut für Organische Chemie und Biochemie, Albert-Ludwigs-Universität, Freiburg im Breisgau, Germany.
Schmidt
U
U
Ziegler
G A
GA
Schweiger
S
S
Hanukoglu
I
I
Schulz
G E
GE
D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005287:Ferredoxin-NADP Reductase / Q000737:chemistry / Q000235:genetics*; D018832:Molecular Chaperones / Q000378:metabolism*; D016296:Mutagenesis