E Chromacin-like peptide in leeches

Neuroendocrinol Lett 2003; 24(3/4):227–232 pii: NEL243403A13 Copyright © Neuroendocrinology Letters www.nel.edu Abstract We demonstrate the presence in leech hemolymph of high levels of a peptide rec-ognized by antiserum directed against bovine chromacin. The purifi cation of the chromacin-like peptide was carried out by an acidic extraction, followed by solid phase and high pressure gel permeation chromatography and reversed-phase HPLC purifi cation. Its sequence (GDFELPSIADPQATFESQRGPSAQQVDK) was established by a combination of techniques, including automated Edman degradation, MALDI-TOF measurement and DOT immunobinding assays with anti-chromogranin A. Mass spectrometry measurement revealed a m/z 3177Da, revealing the fact that the molecule is phosphorylated. ELISA titrations per-formed at each step of the purifi cation revealed a major increase in the level of the peptide (ca. 125 nmol/µl of coelomic fl uid) 15 min after LPS exposure. The increase in chromacin-like peptide levels is both time and concentration depen-dent. The level of this peptide decreased signifi cantly 4 hours after LPS expo-sure. This report is the fi rst discovery of a chromogranin derived like peptide i