Proteolytic processing of the Aplysia egg-laying hormone prohormone

By using matrix-assisted laser desorption/ionization time-of-flight MS, individual peptidergic neurons fromAplysia are assayed. A semiquantitative method is developed for comparing single-cell profiles by using spectral normalization, and peptides are localized to specific cells by mass spectrometric cell mapping. In addition to all previously identified products of the egg-laying hormone (ELH) gene, other peptides are formed from proteolytic hydrolysis of Leu-Leu residues within ELH and acidic peptide (AP). AP exhibits further processing to yield AP1–20 and AP9–27 . These peptides appear to be colocalized in vesicles with ELH, transported to specific neuronal targets, and released in a Ca2+ -dependent manner. A differential peptide distribution is observed at a specific target cell, and a low-frequency variation of AP, [Thr21 ]AP, is detected in a single animal.