Catalytic properties of the human cytochrome P450 2E1 produced by cDNA expression in mammalian cells.
作者： Chris J. Patten , Hiroyuki Ishizaki , Toshifumi Aoyama , Maojung Lee , Shu M. Ning
DOI: 10.1016/0003-9861(92)90258-X
关键词:
摘要: Abstract A full-length cDNA encoding human cytochrome P450 2E1 was expressed in mammalian cell lines using the vaccinia virus expression system. Immunoblot analysis showed that protein reacted with a polyclonal antibody against rat and comigrated from liver microsomes. Hep G2 cells, line which contains both b5 NADPH:P450 oxidoreductase, able to metabolize several known substrates: N-nitrosodimethylamine (NDMA), N-nitrosomethylbenzylamine (NMBzA), p-nitrophenol, phenol, acetaminophen. Apparent Km Vmax values for NDMA demethylation were 22 μ m 173 pmol/min/mg microsomal protein, respectively. TK 143 do not contain b5, displayed of 31 34 Incorporation purified into TK−143 microsomes increased 2.2-fold decreased . Addition resulted 1.6-fold increase Vmax, but no effect on Km. cells shown NMBzA 47 213 protein. lowered 27 , had Vmax. These results demonstrate conclusively is responsible low forms demethylase debenzylase observed these activities are affected by b5.
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