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Mass spectrometry‐based absolute quantification of microsomal cytochrome P450 2D6 in human liver
Author(s) -
Langenfeld Elmar,
Zanger Ulrich M.,
Jung Klaus,
Meyer Helmut E.,
Marcus Katrin
Publication year - 2009
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800680
Subject(s) - cyp2d6 , cytochrome p450 , microsome , human liver , cytochrome , mass spectrometry , chemistry , biology , enzyme , chromatography , biochemistry
We describe the assay of the human cytochrome P450 2D6 in a set of 30 genotyped liver samples using the ‘absolute quantification’ (AQUA) technique. We found approximately 30 fmol CYP2D6 per μg of microsomal protein, with the values spanning from 0 to nearly 80 fmol/μg. This is greater by a factor of 5–10 from the compared to the currently accepted value, which was around 5 fmol/μg. Our results thus suggest that the amount of cytochrome P450 (CYP) enzymes in liver have to be reassessed. We used quantitative Western blotting, calibration standards and activity assays, to validate the results. Our results show, that using the AQUA technique a true assay of CYP2D6 in human liver was possible.