Pertussis Toxin Enhances Proenkephalin Synthesis in Bovine Chromaffin Cells

Abstract: The synthesis of the neuropeptide precursor proenkephalin was measured in bovine adrenal Chromaffin cells following radiolabeling with ³⁵S]methionine. Treatment of Chromaffin cells with pertussis toxin (100 ng/ml) approximately doubled proenkephalin synthesis without altering total protein synthesis. Pertussis toxin pretreat-ment also increased proenkephalin synthesis in chromaf-fin cells exposed to vasoactive intestinal peptide (VIP) and 3-isobutyl-1 -methylxanthine (IBMX). Combinations of IBMX plus nicotine, VIP, or histamine also synergistically enhanced proenkephalin synthesis, with no further elevation when the cells were also pretreated with pertussis toxin. The action of forskolin, a direct activator of adenyl-ate cyclase, on proenkephalin synthesis was similarly potentiated by pertussis toxin or IBMX, presumably reflecting the abilities of both the toxin and this phosphodiester-ase inhibitor to enhance the cyclic AMP response to forskolin. In contrast, increased synthesis of proenkephalin in response to phorbol esters was not affected by pertussis toxin treatment. These results suggest that pertussis toxin potentiates proenkephalin synthesis primarily through inactivation of guanine nucleotide-binding proteins that inhibit adenylate cyclase, although other signaling pathways may also be involved.