Detection of cytochrome P4501A in several species using antibody against a synthetic peptide derived from rainbow trout cytochrome P4501A1
- Univ. of British Columbia, Vancouver, British Columbia (Canada). Faculty of Pharmaceutical Sciences
- Dept. of Fisheries and Oceans, Sidney, British Columbia (Canada). Inst. of Ocean Sciences
Induction of cytochrome P4501A (CYP1A) is being used increasingly as a biomarker to indicate exposure of organisms to environmental contaminants such as some polycyclic and polychlorinated aromatic hydrocarbons. Measurement of CYPIA protein in wildlife would be facilitated by the use of a specific antibody that recognized the isozyme in several species. In the present study, a polyclonal antibody targeted to CYP1A1 was generated using a synthetic peptide corresponding to amino acids 277--294 of the trout enzyme as the antigen of immunization. Specificity of the resulting antibody was assessed by noncompetitive enzyme-linked immunosorbent assay with several purified rat CYP isozymes and by immunoblot analysis with liver microsomes from diverse species. The antibody reacted strongly with the immunizing peptide and with purified rat cytochrome P4501A1 but did not react with rat CYP1A2, a closely related isozyme, or with six other purified rat CYP proteins in enzyme-linked immunosorbent assay. On immunoblots, the antibody recognized a single protein band in hepatic microsomes from the various mammal and fish species tested. Two protein bands were detected in liver microsomes from 3-methylcholanthrene-treated chickens. The results suggest that the antigenic determinant to which the antibody binds is unique to CYP1A and is conserved in different species. Because of its specificity, this anti-peptide antibody should be suitable as a probe to measure CYP1A protein levels in wildlife.
- Sponsoring Organization:
- USDOE
- OSTI ID:
- 675460
- Journal Information:
- Environmental Toxicology and Chemistry, Journal Name: Environmental Toxicology and Chemistry Journal Issue: 3 Vol. 17; ISSN 0730-7268; ISSN ETOCDK
- Country of Publication:
- United States
- Language:
- English
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