title: Atomic force microscopy detection of molecular complexes in multiprotein P450cam containing monooxygenase system.
abstract: 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.
authors.affiliation: Institute of Biomedical Chemistry, Moscow, Russia.
authors.forename: Vadim Yu
authors.initials: VY
authors.lastname: Kuznetsov
authors.forename: Yuri D
authors.initials: YD
authors.lastname: Ivanov
authors.forename: Victor A
authors.initials: VA
authors.lastname: Bykov
authors.forename: Sergey A
authors.initials: SA
authors.lastname: Saunin
authors.forename: Igor A
authors.initials: IA
authors.lastname: Fedorov
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Lemeshko
authors.forename: Hui Bon
authors.initials: HB
authors.lastname: Hoa
authors.forename: Alexander I
authors.initials: AI
authors.lastname: Archakov
mesh_terms: 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
chemical_list: D005288:Ferredoxins; D009097:Multienzyme Complexes; C100137:putidaredoxin; D019475:Camphor 5-Monooxygenase; D009247:NADH, NADPH Oxidoreductases; C022486:putidaredoxin reductase