Chromacin-like peptide in leeches.
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/microl 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.
Salzet
Michel
M
Laboratoire de Neuroimmunologie des Annélides, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq Cedex, France. michel.salzet@univ-lille1.fr
Stefano
George
G
D000595:Amino Acid Sequence; D000818:Animals; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D006458:Hemolymph / Q000378:metabolism; D007865:Leeches / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D010455:Peptides / Q000378:metabolism*; D011863:Radioimmunoassay; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D008070:Lipopolysaccharides; D010455:Peptides