Anti-peptide antibodies to the P4502D subfamily in rat, dog and man

Xenobiotica. 1995 Nov;25(10):1103-9. doi: 10.3109/00498259509061910.


1. In order to obtain specific antibodies of the P4502D subfamily, we generated two anti-peptide antibodies against synthetic peptides, DPAQPPRD (peptide A) and DPTQPPRH (peptide B). The sequence of peptide A occurs in rat P4502D2, P4502D4 and human P4502D6, whereas the sequence of peptide B occurs in the dog P4502D subfamily. These sequences are closely related to an epitope of liver/kidney microsomal autoimmune hepatitis. 2. In immunoblotting studies, the anti-peptide antibody against peptide A recognized a 49-KDa protein in microsomes derived from human lymphoblasts expressing P4502D6 and rat liver. It showed no crossreactivity with microsomes from dog liver. In contrast, the anti-peptide antibody against peptide B recognized a 49-KDa protein only in microsomes of dog liver. These indicate that each anti-peptide antibody has the specificity for the respective sequences of the members of P4502D subfamily, with the species investigated herein. 3. In immunoinhibition studies, the anti-peptide antibodies against peptide B inhibited bunitrolol 4-hydroxylation and propranolol 4,5-hydroxylation, which are mediated by the dog P4502D subfamily. These data suggest that the anti-peptide antibodies against peptide B bind to the native and denatured forms of the P4502D subfamily. 4. The present study has demonstrated that the anti-peptide antibodies against this region are useful for studying the members of the P4502D subfamily.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adrenergic beta-Antagonists / metabolism
  • Amino Acid Sequence
  • Animals
  • Antibodies / immunology*
  • Blotting, Western
  • Cytochrome P-450 CYP2D6
  • Cytochrome P-450 Enzyme System / immunology*
  • Cytochrome P-450 Enzyme System / metabolism
  • Dogs
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunoglobulin G / biosynthesis
  • Immunoglobulin G / immunology
  • In Vitro Techniques
  • Lymphocytes / enzymology
  • Microsomes, Liver / enzymology
  • Mixed Function Oxygenases / immunology*
  • Mixed Function Oxygenases / metabolism
  • Molecular Sequence Data
  • Peptides / chemical synthesis
  • Peptides / immunology*
  • Propanolamines / metabolism
  • Propranolol / metabolism
  • Rats


  • Adrenergic beta-Antagonists
  • Antibodies
  • Immunoglobulin G
  • Peptides
  • Propanolamines
  • Cytochrome P-450 Enzyme System
  • Propranolol
  • Mixed Function Oxygenases
  • Cytochrome P-450 CYP2D6
  • bunitrolol