Premium
Inhibition of Human Cytochrome P450 2D6 by Schering 66712
Author(s) -
Butler Brendan Francis,
Palamanda Jairam,
Nomeir Amin A.,
Guengerich F. Peter,
Furge Laura Lowe
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a670-a
Subject(s) - microsome , recombinant dna , heme , cytochrome p450 , chemistry , enzyme , microbiology and biotechnology , biochemistry , biology , gene
Schering 66712 ( SCH66712 ) is a known mechanism‐based inhibitor of human cytochrome P450 2D6. Spectral binding assays with purified recombinant protein indicate that SCH66712 displays classical Type I binding with ferric P450 2D6 with K S of 0.39± 0.10 μM. Incubation of SCH66712 with purified 2D6 and NADPH resulted in a loss of P450 heme‐CO difference spectrum. Pyridine hemochrome assays also showed loss of heme over time. SDS‐PAGE analysis of inactivation assays with human liver microsomes, recombinant microsomes or reconstituted purified 2D6 demonstrated that 3H‐ and 14C‐labeled SCH66712 were irreversibly bound to the apoprotein in SCH66712 inactivated samples and that the association was NADPH dependent. ESI‐LC‐MS analysis of the SCH66712 inactivated samples showed that the mass of the inactivated 2D6 apoprotein increased by approximately 380 Da ( SCH66712 , 338 Da). (Supported in part by: Howard Hughes Medical Institute grant to Kalamazoo College, Michigan Economic Development Corporation, Dreyfus Foundation).