Assessment of P450 induction in the marmoset monkey using targeted anti-peptide antibodies.
The identity and expression of hepatic P450 enzymes in marmosets was investigated using a panel of anti-peptide antibodies originally targeted against human P450 enzymes. In immunoblotting, of 12 antibodies examined, 10 bound specifically to bands in marmoset liver microsomal fraction corresponding to P450 enzymes. It is proposed that these represent marmoset CYP1A1, CYP1A2, CYP2A, CYP2B, CYP2C forms (CYP2C-1 and CYP2C-2), CYP2D19, CYP3A21 and another CYP3A form (CYP3A-m). The antibodies, together with an anti-marmoset CYP2E1 antibody, were used to investigate the expression of 10 P450 enzymes in marmosets treated with P450-inducing chemicals. Treatment with phenobarbitone caused CYP2B, CYP2C-2 and CYP3A21 levels to increase, rifampicin caused increases in CYP2B and CYP2C-1 and a decrease in CYP3A21 levels, whereas dioxin caused CYP1A1 and CYP1A2 levels to increase and CYP2E1 levels to decrease. Clofibric acid did not induce any P450. P450 enzyme activities were assessed using 8 different substrates and increases were found after treatment with phenobarbitone, rifampicin, and dioxin. However, due to species differences in substrate selectivity, it proved difficult to ascribe these changes to individual P450 enzymes. Thus, the use of anti-peptide antibodies provides a more informative way of assessing the levels of specific P450 enzymes than enzyme activity measurements.
Section on Clinical Pharmacology, Division of Medicine, Imperial College School of Medicine, Hammersmith Hospital, Du Cane Road, London W12 0NN, UK.
T G
TG
Schulz
R
R
Thiel
D
D
Neubert
P J
PJ
Brassil
T
T
Schulz-Utermoehl
A R
AR
Boobis
R J
RJ
Edwards
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002144:Callithrix; D003577:Cytochrome P-450 Enzyme System / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007527:Isoenzymes / Q000096:biosynthesis / Q000737:chemistry; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D010455:Peptides / Q000276:immunology; D000072317:Polychlorinated Dibenzodioxins; D013045:Species Specificity
D000906:Antibodies; D007527:Isoenzymes; D010455:Peptides; D000072317:Polychlorinated Dibenzodioxins; D003577:Cytochrome P-450 Enzyme System