Kuznetsov Vadim Yu, Ivanov Yuri D, Bykov Victor A, Saunin Sergey A, Fedorov Igor A, Lemeshko Sergey V, Hoa Hui Bon, Archakov Alexander I
Institute of Biomedical Chemistry, Moscow, Russia.
Proteomics. 2002 Dec;2(12):1699-705. doi: 10.1002/1615-9861(200212)2:12<1699::AID-PROT1699>3.0.CO;2-R.
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.
原子力显微镜（AFM）技术在蛋白质组学研究中的应用得到了证实，该技术可用于识别和可视化含细胞色素P450cam单加氧酶系统中的单个分子和分子复合物。该方法能够区分二元蛋白质复合物和相应的单体蛋白质，也能区分二元复合物和三元复合物。在云母载体上获得了含细胞色素P450cam单加氧酶系统的组分——细胞色素P450cam（P450cam）、恶臭假单胞菌铁氧还蛋白（Pd）和恶臭假单胞菌铁氧还蛋白还原酶（PdR）——的AFM图像。发现P450cam、Pd和PdR分子的典型高度分别为2.6±0.3 nm、2.0±0.3 nm和2.8±0.3 nm。二元Pd/PdR和P450cam/PdR复合物的测量高度分别为4.9±0.3 nm和5.1±0.3 nm。发现二元P450cam/Pd复合物的典型高度约为（3.9 / 5.7 nm），三元PdR/Pd/P450cam复合物的典型高度约为9.1±0.3 nm。
