Antisera against Periplaneta americana Cu,Zn-superoxide dismutase (SOD): separation of the neurohormone bursicon from SOD, and immunodetection of SODin the central nervous system

In an effort to characterize the insect molting hormone bursicon from the c ockroach, Periplaneta americana, amino acid sequences with high identity to Cu,Zn-superoxide dismutase (SOD) of Drosophila virilis were identified. An tisera against a conserved region of SOD, and a sequence unique to Periplan eta SOD were produced and used to test whether bursicon might be a form of SOD. Western blots of one- and two-dimensional gels revealed that the dimer ic form of SOD and bursicon have a similar molecular mass (30 kDa). The two proteins can be separated. however, according to their different isoelectr ic points. Bursicon is identified in two-dimensional gels by elution from f our unique spots not labeled by the anti-SOD antisera. In sections of Perip laneta nerve cords the antisera labeled glial material surrounding neuronal somata close to the neural sheath. Bursicon, however, is contained in uniq ue cell pairs in the ganglia of the ventral nerve cord. These neurons were labeled with new antisera produced against novel sequences of one of the fo ur above-mentioned bursicon active spots. The results show unequivocally th at SOD and bursicon are distinctly different proteins. Furthermore, the ant i-SOD antisera provided a tool to isolate and sequence bursicon. (C) 1999 E lsevier Science Ltd. All rights reserved.