Optical biosensor study of ternary complex formation in a cytochrome P4502B4 system.
The optical biosensor method was used for the revelation of ternary complexes, formed by the full-length NADPH-cytochrome P450 reductase (d-Fp) and cytochromes P4502B4 (d-2B4) and b5 (d-b5) in the course of their interactions within the reconstituted d-2B4-containing system. Based on the lack of competition between d-b5 and d-Fp for the binding sites on immobilized 2B4 (3) as well as on the analysis of data obtained in the three proteins' dissociation reactions, the possibility of formation of ternary complexes through the interactions between membranous hydrophobic fragments of proteins was substantiated. All the complexes obtained were productive.
Ivanov
Y D
YD
Institute of Biomedical Chemistry RAMS, Moscow, Russia. yuiv@ibmh.msk.su
Kanaeva
I P
IP
Archakov
A I
AI
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases; D001667:Binding, Competitive; D015374:Biosensing Techniques; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000378:metabolism*; D015786:Cytochromes b5 / Q000737:chemistry / Q000378:metabolism; D004800:Enzymes, Immobilized; D006900:Hydroxylation; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D009251:NADPH-Ferrihemoprotein Reductase / Q000737:chemistry / Q000378:metabolism; D055095:Optics and Photonics; D011485:Protein Binding; D011817:Rabbits; D013250:Steroid Hydroxylases / Q000737:chemistry* / Q000378:metabolism*
D004800:Enzymes, Immobilized; D046911:Macromolecular Substances; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; C050356:steroid 15-alpha-hydroxylase; D009251:NADPH-Ferrihemoprotein Reductase