caa a22 4500 477117155 CHVBK 20180405111625.0 cr unu---uuuuu 170330e19960801xx s 000 0 eng 10.1007/BF02112359 doi (NATIONALLICENCE)springer-10.1007/BF02112359 Immunochemical relationships of cytochrome P4503A-like proteins in teleost fish [Elektronische Daten] [Malin Celander, Donald Buhler, Lars Förlin, Anders Goksøyr, Cristobal Miranda, Bruce Woodin, John Stegeman] Multiple P450 proteins have been purified from several teleost species, including rainbow trout (Oncorhynchus mykiss), scup (Stenotomus chrysops) and Atlantic cod (Gadus morhua). Identity, relationships and/or functions have been established in these fish species for the cytochrome P4501 As. Information about the structure, function, regulation and relationships of other piscine cytochrome P450 (CYP) proteins is sparse. In the present study we have focused on constitutively expressed CYP forms, P450con and LMC5 isolated from rainbow trout, P450A from scup, and P450b from Atlantic cod, and we consider evidence for the relationship of these proteins to mammalian members of the CYP3A subfamily. Reciprocal western blot analysis shows that P450con and LMC5, isolated from rainbow trout in two different laboratories, are closely related and ostensibly identical proteins. These trout proteins show specific reciprocal cross-reactivity with scup P450A, and polyclonal antibodies (PAb) to the trout and scup proteins both recognize cod P450b, indicating that rainbow trout P450con/LMC5, scup P450A and cod P450b are immunochemically-related proteins. In analyses of liver microsomes of trout, scup and cod, PAb to trout P450con/LMC5 and scup P450A recognize only bands that are identical in migration to the CYP proteins purified from these species, and which were used as immunogens. These CYP proteins purified from fish are each immunochemically-related to mammalian CYP3A proteins, showing recognition by PAb to human CYP3A4 and to rat CYP3A1. PAb to the mammalian CYP3As also recognize the same bands in liver microsomes from these fish species as seen by PAb to the fish proteins. These results strongly suggest that these fish proteins are members of theCYP3 gene family and probably theCYP3A subfamily. Although sequence analysis is required before their designation in the CYP3A subfamily can be confirmed and specified, we refer to these as CYP3A-like. Immunoblot analyses of hepatic microsomes from other fish species with PAb to scup P450A and trout P450con show that multiple CYP3A-like proteins are expressed in liver of several species, including killifish (Fundulus heteroclitus) and winter flounder (Pleuronectes americanus). Important questions still remain to be addressed concerning CYP3A structure, multiplicity, physiological function, regulation and metabolism of endogenous as well as exogenous substrates in fish. Kugler Publication bv Amsterdam, 1996 cytochrome P450 nationallicence CYP3A nationallicence fish nationallicence immunoblotting nationallicence Celander Malin Biology Department, Woods Hole Oceanographic Institution, 02543, Woods Hole, MA, USA aut Buhler Donald Department of Agricultural Chemistry and Marine Freshwater Biomedical Center, Oregon State University, 97331, Corvallis, OR, USA aut Förlin Lars Department of Zoophysiology, University of Göteborg, Medicinaregatan 18, S-413 90, Göteborg, Sweden aut Goksøyr Anders Laboratory of Marine Molecular Biology, University of Bergen, HIB, N-5020, Bergen, Norway aut Miranda Cristobal Department of Agricultural Chemistry and Marine Freshwater Biomedical Center, Oregon State University, 97331, Corvallis, OR, USA aut Woodin Bruce Biology Department, Woods Hole Oceanographic Institution, 02543, Woods Hole, MA, USA aut Stegeman John Biology Department, Woods Hole Oceanographic Institution, 02543, Woods Hole, MA, USA aut Fish Physiology and Biochemistry Kluwer Academic Publishers 15/4(1996-08-01), 323-332 0920-1742 15:4<323 1996 15 10695 https://doi.org/10.1007/BF02112359 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF02112359 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Celander Malin Biology Department, Woods Hole Oceanographic Institution, 02543, Woods Hole, MA, USA aut NATIONALLICENCE 700 1- Buhler Donald Department of Agricultural Chemistry and Marine Freshwater Biomedical Center, Oregon State University, 97331, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Förlin Lars Department of Zoophysiology, University of Göteborg, Medicinaregatan 18, S-413 90, Göteborg, Sweden aut NATIONALLICENCE 700 1- Goksøyr Anders Laboratory of Marine Molecular Biology, University of Bergen, HIB, N-5020, Bergen, Norway aut NATIONALLICENCE 700 1- Miranda Cristobal Department of Agricultural Chemistry and Marine Freshwater Biomedical Center, Oregon State University, 97331, Corvallis, OR, USA aut NATIONALLICENCE 700 1- Woodin Bruce Biology Department, Woods Hole Oceanographic Institution, 02543, Woods Hole, MA, USA aut NATIONALLICENCE 700 1- Stegeman John Biology Department, Woods Hole Oceanographic Institution, 02543, Woods Hole, MA, USA aut NATIONALLICENCE 773 0- Fish Physiology and Biochemistry Kluwer Academic Publishers 15/4(1996-08-01), 323-332 0920-1742 15:4<323 1996 15 10695 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer