Evidence that rabbit cytochrome P-450 K is encoded by the plasmid pP-450 PBc2.
The complete coding sequence for P-450 PBc2 was inserted into a T7-phage promoter system, and a capped cRNA was generated using T7 RNA polymerase. The P-450 PBc2 cRNA was translated in a rabbit reticulocyte lysate. The in vitro translation product was indirectly immunoprecipitated by the monoclonal antibodies 2F5 and 3C3 that recognize P-450 K. SDS-polyacrylamide gel electrophoresis revealed that the translated protein product exhibits the same relative electrophoretic mobility as P-450 K. The N-terminal amino acid sequence was determined to be MDLVVVLGL-LS-LLLLSL- for P-450 K immunopurified from rabbit kidney using the monoclonal antibody 2F5. This sequence agrees with the predicted amino acid sequence derived from the P-450 PBc2 cDNA. These results indicate that P-450 K or a protein closely related to P-450 K is encoded by the plasmid pP-450 PBc2.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000276:immunology; D007668:Kidney / Q000201:enzymology*; D010957:Plasmids; D014176:Protein Biosynthesis; D011817:Rabbits
D000911:Antibodies, Monoclonal; D003577:Cytochrome P-450 Enzyme System