Atomic force microscopy detection of molecular complexes in multiprotein P450cam containing monooxygenase system.
The application of atomic force microscopy (AFM) technique in proteomic research, identification and visualization of individual molecules and molecular complexes within the P450cam containing monooxygenase system was demonstrated. The method distinguishes between the binary protein complexes and appropriate monomeric proteins and, also, between the binary and ternary complexes. The AFM images of the components of a cytochrome P450cam containing monooxygenase system - cytochrome P450cam (P450cam), putidaredoxin (Pd) and putidaredoxin reductase (PdR) - were obtained on a mica support. The molecules of P450cam, Pd and PdR were found to have typical heights of 2.6 +/- 0.3 nm, 2.0 +/- 0.3 and 2.8 +/- 0.3 nm, respectively. The measured heights of the binary Pd/PdR and P450cam/PdR complexes were 4.9 +/- 0.3 nm and 5.1 +/- 0.3 nm, respectively. The binary P450cam/Pd complexes were found to have a typical height of about (3.9 / 5.7 nm) and the ternary PdR/Pd/P450cam complexes, a typical height of about 9.1 +/- 0.3 nm.
Kuznetsov
Vadim Yu
VY
Institute of Biomedical Chemistry, Moscow, Russia.
Ivanov
Yuri D
YD
Bykov
Victor A
VA
Saunin
Sergey A
SA
Fedorov
Igor A
IA
Lemeshko
Sergey V
SV
Hoa
Hui Bon
HB
Archakov
Alexander I
AI
D019475:Camphor 5-Monooxygenase / Q000737:chemistry* / Q000648:ultrastructure; D005288:Ferredoxins / Q000737:chemistry / Q000648:ultrastructure; D018625:Microscopy, Atomic Force; D009097:Multienzyme Complexes; D009247:NADH, NADPH Oxidoreductases / Q000737:chemistry / Q000648:ultrastructure; D010084:Oxidation-Reduction
D005288:Ferredoxins; D009097:Multienzyme Complexes; C100137:putidaredoxin; D019475:Camphor 5-Monooxygenase; D009247:NADH, NADPH Oxidoreductases; C022486:putidaredoxin reductase