RT Journal Article
SR Electronic
T1 An investigation of the antigenic determinants on chloroperoxidase and purified rat liver microsomal cytochrome P-450b.
JF Drug Metabolism and Disposition
JO Drug Metab Dispos
FD American Society for Pharmacology and Experimental Therapeutics
SP 518
OP 523
VO 15
IS 4
A1 Pandey, R N
A1 Kuemmerle, S C
A1 Hollenberg, P F
YR 1987
UL http://dmd.aspetjournals.org/content/15/4/518.abstract
AB Chloroperoxidase (CPO) exhibits many physicochemical and catalytic properties similar to those of the bacterial and microsomal cytochromes P-450. Therefore, the possible similarities between the antigenic determinants of CPO and rat liver microsomal cytochrome P-450b were investigated. Polyclonal antibodies against CPO and rat liver cytochrome P-450b were raised in rabbits and used to investigate the antigenic cross-reactivity between CPO and P-450b. Although anti-CPO antibodies were capable of inhibiting the ethyl hydroperoxide-supported N,N-dimethylaniline (DMA) demethylation activity of CPO by more than 80%, they were unable to inhibit the NADPH-supported demethylation of DMA by cytochrome P-450b in the reconstituted system. The ethyl hydroperoxide-supported demethylation of DMA by CPO was not affected by the addition of anti-P-450b antibodies which inhibited cytochrome P-450 activity greater than 90%. In order to probe for the possible existence of common antigenic determinants which were not involved in catalytic activity, the cross-reactivities were investigated using enzyme-linked immunosorbent assays. There was no cross-reactivity between anti-CPO and cytochrome P-450b, or anti-P-450b and CPO using enzyme-linked immunosorbent assays. When control, phenobarbital-, isosafrole-, and beta-naphthoflavone-induced rat and rabbit liver microsomes and CPO were analyzed by Western blotting and developed with anti-P-450 antibodies, only the phenobarbital- and isosafrole-induced microsomes showed a positive reaction in the P-450 region. When anti-CPO antibodies were used on Western blots of the same series of proteins, a positive reaction was observed only with CPO.(ABSTRACT TRUNCATED AT 250 WORDS)