CROSS-REACTIVITY OF AN ANTIBODY TO SPINY LOBSTER P450 2L WITH MICROSOMES FROM OTHER SPECIES

The gene for a new form of P450, CYP2L, was recently sequenced in this laboratory, using a primer designed from the N-terminal amino acid se quence of a P450 partially purified from spiny lobster hepatopancreas microsomes. The partially purified (75% pure) P450 was used to immuniz e rabbits. Serum containing the rabbit anti-CYP2L antibody was demonst rated to possess good cross-reactivity on Western blots with microsoma l preparations from hepatopancreas of the spiny lobster. In Western bl ot studies with microsomal preparations from other aquatic invertebrat es, the horseshoe crab, the blue crab, the slipper lobster, the Americ an lobster and the brown shrimp, the slipper lobster microsomal prepar ations showed strong cross-reactivity with the spiny lobster antibody and the American lobster weak cross-reactivity. Hepatic microsomes fro m all of the vertebrate species studied showed cross-reactivity with t he anti-CYP2L antibody. The fish species studied were killifish, clear -nose skate, snook and catfish. Strong cross-reactivity was found with killifish and catfish, and weak cross-reactivity was found with skate and snook in the 45-66 kDa region. Cross-reactivity was also found wi th microsomes from male control and phenobarbital-induced rats. These studies suggest that epitopes in CYP2L are also present in other inver tebrate and vertebrate species. Copyright (C) 1996 Elsevier Science Lt d