Immuno-detection of proteins by cytochrome P450 antibodies in five species of sea anemones

The presence and nature of immunoreactive proteins to several cytochrome P450 (CYP) antibodies in the columnar or whole body microsomal fraction of five sea anemone species (Anthopleura elegantissima, Aiptasia pallida, Anthopleura xanthogrammica, Bunodosoma cavernata and Condylactis gigantea) were probed by Western blot analysis. Anthopleura elegantissima, A. xanthogrammica, B. cavernata and C. gigantea columnar microsomes contained proteins that crossreacted with monoclonal anti-scup CYP 1A1 (˜ 70 kDa), polyclonal anti-trout CYP 2K1 (˜ 40 kDa) and polyclonal anti-trout CYP 3A1 (˜ 30 kDa). Aiptasia pallida whole body microsomes contained various proteins that cross-reacted with scup CYP 1A1, trout CYP 2K1 and trout CYP 3A1 antibodies. 3-Methylcholantrene (3MC, 20 mg/kg) did not induce proteins recognized by anti-CYP 1A1 in A. elegantissima. Our results indicate the presence in anemone microsomes of various proteins with epitope regions recognized by several CYP antibodies; the most strongly recognized proteins differ from the 50–60 kDa molecular weights which are typically ascribed to P450s of vertebrate microsomes. In all anemones, proteins in the 50–60 kDa range are very weakly recognized even at relatively high protein concentrations.