Hamster hepatic cytochrome b5: purifications, immunochemical properties, and in vitro synthesis.
Cytochrome b5 has been purified from hamster liver microsomes. Both Ouchterlony double-diffusion and rocket immunoelectrophoresis experiments indicate that no immuno-cross-reactivity exists between guinea-pig anti-rabbit cytochrome b5 antibody and hamster cytochrome b5. However, anti-rabbit b5 IgG inhibited both hamster microsomal NADH-cytochrome c reductase and NADPH-dependent 7-ethoxycoumarin-O-deethylase activities. Hamster cytochrome b5 stimulated several reconstituted hamster cytochrome P-450-dependent monooxygenase activities and this stimulatory effect could be inhibited by antibody against rabbit cytochrome b5. Two-dimensional iodinated tryptic peptide mapping experiments provided evidence that the polypeptide fingerprint of hamster cytochrome b5 is substantially different from the fingerprints of cytochrome b5 isolated from rabbit, rat and bovine. We also studied the in vitro synthesis of hamster cytochrome b5 from liver mRNA using a wheat germ lysate system. A 16 kDa polypeptide, which is the same size as hamster cytochrome b5, was immunoprecipitated by antibody against rabbit b5. This experiment suggested that in vitro synthesized hamster cytochrome b5 is recognized by a heterologous antibody. Thus, hamster and rabbit cytochrome b5 do share some common immuno-determinants which may be located close to the heme-binding active site.
D000818:Animals; D006224:Cricetinae; D003429:Cross Reactions; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D003573:Cytochrome b Group / Q000276:immunology / Q000302:isolation & purification*; D015786:Cytochromes b5; D004591:Electrophoresis, Polyacrylamide Gel; D007122:Immunoelectrophoresis; D008862:Microsomes, Liver / Q000032:analysis*; D010455:Peptides / Q000032:analysis; D012333:RNA, Messenger; D011817:Rabbits / Q000276:immunology; D014158:Transcription, Genetic
D003573:Cytochrome b Group; D010455:Peptides; D012333:RNA, Messenger; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System