PMID: 8946469Oct 1, 1996Paper

Selective expression of CYP3A5 and not CYP3A4 in human blood

Pharmacogenetics
S K JanardanP B Watkins

Abstract

There is a marked variation between people in the activity of CYP3A4 in liver and intestine. We reasoned that if CYP3A4 was expressed in peripheral blood cells, a simple blood based test of CYP3A4 phenotype might be feasible. We prepared peripheral blood smears from healthy volunteers and performed immunostaining with a rabbit polyclonal antibody that selectively reacts with enzymes within the CYP3A subfamily. Staining was observed only within the cytoplasm of neutrophils (PMNs). cDNA prepared from isolated PMNs and mononuclear cells was subjected the polymerase chain reaction using as primers synthetic oligonucleotides that selectively amplify fragments of each known CYP3A cDNAs (CYP3A3, CYP3A4, CYP3A5, and CYP3A7). Amplification was only obtained with the CYP3A5 specific oligonucleotides, predominantly in PMNs, and the identity of the amplified fragment was confirmed by sequencing. Next, whole white cell homogenate prepared from human blood was reacted on immunoblots with a monoclonal antibody that recognizes all CYP3A proteins or an absorbed polyclonal antibody that recognizes only CYP3A5. Both antibodies recognized a protein in the white cells that comigrated with purified CYP3A5. However, metabolism of midazolam, a substra...Continue Reading

Citations

Oct 31, 2002·Advanced Drug Delivery Reviews·Jatinder K LambaKenneth E Thummel
May 3, 2003·Regulatory Toxicology and Pharmacology : RTP·Ulrike BernauerUrsula Gundert-Remy
Aug 13, 1999·Experimental and Toxicologic Pathology : Official Journal of the Gesellschaft Für Toxikologische Pathologie·H RaunioR J Edwards
Oct 13, 2006·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Su-Jun LeeRon H N van Schaik
Mar 27, 2002·Biological & Pharmaceutical Bulletin·Fusako NagaiHiro-Omi Tamura
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Jul 16, 2011·Drug Metabolism and Drug Interactions·Werner Cordier, Vanessa Steenkamp
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Sep 14, 2002·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Ina KochLeszek Wojnowski
Dec 18, 2001·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Ali AsgharStephen D Hall
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