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NEUROENDOCRINOLOGY LETTERS
including Psychoneuroimmunology, Neuropsychopharmacology,
Reproductive Medicine, Chronobiology
and Human Ethology, ISSN 0172–780X

NEL Vol.24 Nos.3/4, Jun-Aug 2003

ORIGINAL ARTICLE

Chromacin-like peptide in leeches

2003; 24:227232
pii: NEL243403A13
PMID: 14523362

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Chromacin-like peptide in leeches

Michel Salzet 1 & George Stefano 2

1. Laboratoire de Neuroimmunologie des Annélides, Université des Sciences et Technologies de Lille, Villeneuve d’Ascq Cedex, France.

2. Neuroscience Research Institute, State University of New York College at Old Westbury, Old Westbury, NY 11568-0210, USA.

Submitted: November 6, 2002
Accepted: December 16, 2002

Key words:
neuroimmunity, chromacin-like peptide, invertebrate, lophotrochozoans, Annelids

 

Abstract

We demonstrate the presence in leech hemolymph of high levels of a peptide recognized by antiserum directed against bovine chromacin. The purification 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 purification. 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 performed at each step of the purification revealed a major increase in the level of the peptide (ca. 125 nmol/l of coelomic fluid) 15 min after LPS exposure. The increase in chromacin-like peptide levels is both time and concentration dependent. The level of this peptide decreased significantly 4 hours after LPS exposure. This report is the first discovery of a chromogranin derived like peptide in invertebrates.

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