Isolation, structural characterization and biological function of a lysine-conopressin in the central nervous system of the Pharyngobdellid leech Erpobdella octoculata

Abstract

Several neuropeptides are suspected to act on the control of hydric balance in leeches. One of these peptides, a peptide immunoreactive to an antibody against oxytocin, was previously characterized from the central nervous system of the leech Erpobdella octoculata [Salzet, M., Wattez, C., Verger-Bocquet, M., Beauvillain, J.-C. & Malecha, J. (1993) Brain Res. 601, 173–184].

This paper reports the isolation from the central nervous system of E. octoculata of another peptide of the oxytocin/vasopressin family; a lysine-vasopressin-like molecule. Its purification was performed by reverse-phase high-performance liquid chromatography combined with both dot immunobinding assay and enzyme-linked immunosorbent assay for lysine-vasopressin. The amino acid sequence was established by Edman degradation and confirmed by electrospray-mass-spectrometry measurement. The nonapeptide obtained corresponded to the lysine-conopressin previously isolated from the venom of the mollusc Conus geographus [Cruz, L. L., de Santos, V., Zafaralla, G. C., Ramilo, C. A., Zeikus, R., Gray, W. R. & Olivera, B. M. (1987) J. Biol. Chem. 262, 15821–15824]. In leeches, synthetic lysine-conopressin exerts a diuretic effect which can be compared to that of the arginine-vasopressin-like peptide isolated in the Insect Locusta migratoria [Proux, J., Miller, C. A., Li, J. P., Carney, R. L., Girardie, A., Delaage, M. & Schooley, D. A. (1987) Biochem. Biophys. Res. Commun. 149, 180–186].

Abbreviations

DIA

dot immunobinding assay

RT

retention time