Neuropeptide processing activity in Aplysia bag cell homogenates.

The neurosecretory bag cells of the mollusk, Aplysia, generate a peptide egg-laying hormone (ELH) from a 29,000 Dalton precursor protein by proteolytic cleavage to a 6-9,000 Dalton intermediate, followed by cleavage of the intermediate. We report here the initial characterization of these cleavage activities. Homogenates of bag cells in low ionic strength buffer process endogenous precursor to a peptide which is indistinguishable from ELH in molecular weight and isoelectric point. Non-specific proteolysis in the homogenates is not detectable. The pH optimum for cleavage of the precursor and the intermediate is 5.5-6.5. The cleavage activities exhibit a substantial degree of membrane association, and the inhibitor profile of each is characteristic of a thiol protease without a metal cofactor requirement. Precursor cleavage activity differs from that of the intermediate cleaving activity in inhibitor profile, solubility, and slightly, in pH optimum.