|Title||[A covalently linked complex of cytochrome P-450 with adrenodoxin. Localization of the adrenodoxin binding site of cytochrome P-450].|
|Publication Type||Journal Article|
|Year of Publication||1988|
|Authors||Turko, IV, Adamovich, TB, Kirillova, NM, Usanov, SA, Chashchin, VL|
|Date Published||1988 Nov|
|Keywords||Adrenodoxin, Amino Acid Sequence, Binding Sites, Cross-Linking Reagents, Cytochrome P-450 Enzyme System, Molecular Sequence Data|
Cytochrome P-450scc and adrenodoxin were cross-linked with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide. The sample containing 94% of a cross-linked complex and 6% of free cytochrome P-450scc was obtained after purification on cholate-Sepharose. Cytochrome P-450scc in the cross-linked complex is not reduced in the presence of NADPH and adrenodoxin reductase, but completely preserves its high spin form in the presence of Tween-20 or pregnenolone. The use of radioactive labelled adrenodoxin, chemical cleavage of cytochrome P-450scc from the cross-linked complex by o-iodosobenzoic acid and HPLC for separation of peptides demonstrated that the cytochrome P-450scc complex with adrenodoxin was cross-linked through two amino acid sequences of cytochrome P-450scc, i.e., Leu 88-Trp108 and Leu368-Trp417.