Atomic force microscopy revelation of molecular complexes in the multiprotein cytochrome P450 2B4-containing system.
The application of atomic force microscopy (AFM) to the identification and visualization of individual molecules and their complexes in a reconstituted monooxygenase P450 2B4 system without the phospholipid was demonstrated. The method employed in this study distinguishes the monomeric proteins from their binary complexes and, also, the binary from the ternary complexes. The AFM images of the full-length P450 2B4 system's constituent components - cytochrome P450 2B4 (2B4), NADPH-cytochrome P450 reductase and cytochrome b5 (b5), were obtained on highly-oriented pyrolitic graphite. The typical heights of the d-2B4, d-flavoprotein (Fp) and d-b5 molecules were measured and found to be 2.2 +/- 0.2, 2.3 +/- 0.2 and 1.8 +/- 0.1 nm, respectively. The measured heights of the binary d-Fp/d-2B4 and d-2B4/d-b5 complexes were estimated to be 3.4 +/- 0.2 and 2.8 +/- 0.2 nm, respectively. No formation of d-Fp/d-b5 complexes was registered. The ternary d-Fp/d-2B4/d-b5 complexes were visualized and their heights were found to be roughly equal to 4.3 +/- 0.3 nm and 6.2 +/- 0.3 nm.
V.N.Orekhovich Institute of Biomedical Chemistry RAMS, Moscow, Russia.
Vadim Yu
VY
Kuznetsov
Yuri D
YD
Ivanov
Alexander I
AI
Archakov
D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000737:chemistry*; D000072467:Cytochrome P450 Family 2; D046911:Macromolecular Substances; D018625:Microscopy, Atomic Force; D008862:Microsomes, Liver / Q000737:chemistry; D008970:Molecular Weight; D020836:Protein Structure, Quaternary; D011817:Rabbits
D046911:Macromolecular Substances; D001189:Aryl Hydrocarbon Hydroxylases; D000072467:Cytochrome P450 Family 2; C465241:cytochrome P-450 CYP2B4 (rabbit)