[Synthesis in Escherichia coli and certain properties of hybrid protein consisting of a modified precursor of adrenodoxin reductase and shortened precursor of adrenodoxin].
Some aspects of formation and functioning of the cholesterol hydroxylase system were studied. A hybrid protein was synthesized in E. coli composed of the modified form of the (NADPH)adrenodoxin reductase precursor (N-terminal domain) and the shortened adrenodoxin precursor (C-terminal domain). The modified reductase precursor contained 12 extra amino acid residues at the N-terminus and the N-terminally shortened adrenodoxin precursor had 17 C-terminal amino acids of its targeting presequence. The hybrid reduced cytochrome P450scc in a reconstituted system. Thus, neither the extra 44 amino acids at the N-terminus of the reductase nor the 17 amino acid linker affected the interaction of the active sites in the hybrid protein. These modifications do not interfere with the binding of prosthetic groups and formation of the active sites of two enzymes in the E. coli cells. Modified N-terminal sequence of the hybrid does not affect its import into heterologous mitochondria.
Isaeva
L V
LV
Spiridonova
V A
VA
Liumovich
D
D
Kovaleva
I E
IE
Chernogolov
A A
AA
Usanov
A S
AS
Luzikov
V N
VN
D000302:Adrenal Cortex / Q000201:enzymology; D000325:Adrenodoxin / Q000096:biosynthesis* / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis / Q000235:genetics; D004274:DNA, Recombinant; D004792:Enzyme Precursors / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D005287:Ferredoxin-NADP Reductase / Q000096:biosynthesis* / Q000235:genetics; D008928:Mitochondria / Q000201:enzymology; D008969:Molecular Sequence Data; D010957:Plasmids; D011498:Protein Precursors / Q000096:biosynthesis* / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D004274:DNA, Recombinant; D004792:Enzyme Precursors; D011498:Protein Precursors; D011994:Recombinant Proteins; D000325:Adrenodoxin; D002786:Cholesterol Side-Chain Cleavage Enzyme; D005287:Ferredoxin-NADP Reductase
Sintez v Escherichia coli i nekotorye svoĭstva gibridnogo belka, sostoiashchego iz modifitsirovannogo predshestvennika adrenodoksinreduktazy i ukorochennogo predshestvennika adrenodoksina.