HPLC and electrospray ionization mass spectrometry as tools for the identification of APGWamide-related peptides in gastropod and bivalve mollusks: comparative activities on Mytilus muscles

Abstract

The APGWamide-related neuropeptides, predicted by the cDNA of the APGWamide precursor of Mytilus edulis, have been sought by means of HPLC and electrospray mass ionization. The three predicted peptides KPGWamide, RPGWamide and TPGWamide were detected in the three main muscles and surprisingly an ion at m/z 429 corresponding to the gastropod peptide APGWamide was also demonstrated. Similar investigations performed in Lymnaea stagnalis central nervous system (CNS) revealed the occurrence of mussel APGWamide-related peptides (APGWamide-RPs) demonstrating for the first time the presence and the expression of the two precursors in both gastropod and bivalve mollusks. The absence of homologous domain in the Mytilus precursor [P. Favrel, M. Mathieu, Molecular cloning of a cDNA encoding the precursor of Ala-Pro-Gly-Trp-amide related neuropeptides from the bivalve Mytilus edulis. Neurosci. Lett. 1996;205:210–214] and the Lymnaea precursor [A.B. Smit, C.R. Jiménez, R.W. Dirks, R.P. Croll, W.P.M. Geraerts, Characterization of cDNA clone encoding multiple copies of the neuropeptide APGWamide in the molluscs Lymnaea stagnalis. J. Neurosci. 1992;12:1709–1715] eliminates the hypothesis of an alternative splicing of a single gene and suggests the likelihood of two genes probably resulting from duplication of an ancestral gene before the divergence between gastropods and bivalves. The similar potency observed on contraction assay and the differential distribution of the various peptides suggest that they may exert distinct activities on multiple targets.