Mass spectrometry‐based absolute quantification of microsomal cytochrome P450 2D6 in human liver |
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| Authors: | Elmar Langenfeld Dr Ulrich M Zanger Klaus Jung Helmut E Meyer Katrin Marcus |
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| Institution: | 1. Funktionelle Proteomik, Medizinisches Proteom‐Center, Ruhr‐Universit?t Bochum, Bochum, Germany;2. Dr. Margarete Fischer Bosch‐Institut für klinische Pharmakologie, Stuttgart, Germany;3. Abteilung Medizinische Statistik, Georg‐August‐Universit?t G?ttingen, G?ttingen, Germany;4. Medizinische Proteomik/Bioanalytik, Medizinisches Proteom‐Center, Ruhr‐Universit?t Bochum, Bochum, Germany |
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| Abstract: | 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. |
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| Keywords: | Endoplasmic reticulum Mass spectrometry Membrane proteins Quantitative analysis Western blot analysis |
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