Marmoset CYP3A21, a model for human CYP3A4: protein expression and functional characterization of the promoter.
Due to its small size and the relative evolutionary proximity, the marmoset has been proposed as a model for studies of human drug interactions and metabolism. The current study investigated the expression and regulation of marmoset CYP3A using mass spectrometry and reporter gene techniques. Expression levels of hepatic marmoset CYP3A protein range from 51 to 123 pmol mg-1 total protein (mean 85.2 pmol mg-1, n = 10) and CYP3A21 is the dominant hepatic CYP3A protein in marmosets. The sequence similarity between human CYP3A4 and CYP3A21 across the first 7.5 kb of the cloned CYP3A21 promoter is 88% within the xenobiotic-responsive enhancer module (XREM) and the proximal promoter. Both regulatory modules confer transcriptional activation of CYP3A21-luciferase reporter gene constructs cotransfected with hPXR in intestinal LS174T cells. The pronounced response to rifampin and the moderate response to dexamethasone were similar to those observed with CYP3A4. Taken collectively, these data establish substantial similarities in expression and gene regulation between marmoset CYP3A21 and human CYP3A4. CYP3A21 may be an equivalent of CYP3A4 in New World monkeys, consistent with the phylogenetic relationship between these genes. The marmoset, therefore, appears to be a suitable in vivo model to study CYP3A4 function and regulation.
Koehler
S C
SC
Klinische Chemie, Uni-Klinikum Göttingen, Robert-Koch-Strasse 40, 37075 Göttingen, Germany. koehler@med.uni-goettingen.de
Von Ahsen
N
N
Schlumbohm
C
C
Asif
A R
AR
Goedtel-Armbrust
U
U
Oellerich
M
M
Wojnowski
L
L
Armstrong
V W
VW
D000595:Amino Acid Sequence; D000818:Animals; D001189:Aryl Hydrocarbon Hydroxylases / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D002144:Callithrix / Q000235:genetics* / Q000378:metabolism*; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D004247:DNA / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D008099:Liver / Q000201:enzymology; D008297:Male; D023421:Models, Animal; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012293:Rifampin / Q000494:pharmacology; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D053719:Tandem Mass Spectrometry; D014162:Transfection; D015262:Xenobiotics / Q000378:metabolism
D011994:Recombinant Proteins; D015262:Xenobiotics; D003907:Dexamethasone; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D001189:Aryl Hydrocarbon Hydroxylases; D051544:Cytochrome P-450 CYP3A; C105145:cytochrome P-450 CYP3A21 (marmoset); C510163:CYP3A4 protein, human; D012293:Rifampin