par Baer, Brian B.R.;Schuman, Jason J.T.;Campbell, Patricia A.P.;Cheesman, Matthew M.J.;Nakano, Mariko;Moguilevsky, Nicole 
;Kunze, Kent K.L.;Rettie, Allan A.E.
Référence Biochemistry, 44, 42, page (13914-13920)
Publication Publié, 2005-10
;Kunze, Kent K.L.;Rettie, Allan A.E.Référence Biochemistry, 44, 42, page (13914-13920)
Publication Publié, 2005-10
Article révisé par les pairs
| Résumé : | Typical cytochrome P450s secure the heme prosthetic group with a cysteine thiolate ligand bound to the iron, electrostatic interactions with the heme propionate carboxylates, and hydrophobic interactions with the heme periphery. In addition to these interactions, CYP4B1 covalently binds heme through a monoester link furnished, in part, by a conserved I-helix acid, Glu310. Chromatography, mass spectrometry, and NMR have now been utilized to identify the site of attachment on the heme. Native CYP4B1 covalently binds heme solely at the C-5 methyl position. Unexpectedly, recombinant CYP4B1 from insect cells and Escherichia coli also bound their heme covalently at the C-8 methyl position. Structural heterogeneity may be common among recombinant CYP4 proteins because CYP4A3 exhibited this duality. Attempts to evaluate functional heterogeneity were complicated by the complexity of the system. The phenomenon of covalent heme binding to P450 provides a novel method for assessing microheterogeneity in heme orientation and raises questions about the fidelity of heme incorporation in recombinant systems. © 2005 American Chemical Society. |
