Environmental Biotechnology
EJB Electronic Journal of Biotechnology ISSN: 0717-3458 Vol.4 No. 1, Issue of April 15, 2001
© 2001 by Universidad Católica de Valparaíso -- Chile Received November 17, 2000 / Accepted January 11, 2001
RESEARCH ARTICLE

Purification of a competitive antagonist for calcitonin gene related peptide action from sardine hydrolysates

Marthe Rousseau
Station de Biologie Marine
Muséum National d'Histoire Naturelle
CNRS FRE 2125 BP 225 29182
Concarneau Cedex, France
E-Mail: rousseam@gmx.net

Irineu Batista
Instituto de Investigação das Pescas e do Mar
Avenida de Brasilia 1400
Lisbon, Portugal
Tel: 334-844-2306
Fax: 334-844-4487
E-mail: irineu@ipimar.pt

Yves Le Gal
Station de Biologie Marine
Muséum National d'Histoire Naturelle
CNRS FRE 2125 BP 225 29182
Concarneau Cedex, France
E-mail: ylegal@mnhn.fr

Martine Fouchereau-Peron*
Station de Biologie Marine
Muséum National d'Histoire Naturelle
CNRS FRE 2125 BP 225 29182
Concarneau Cedex, France
Tel: 33 (0) 2 98 97 06 59
Fax: 33 (0) 2 98 97 81 24
E-mail: peron@mnhn.fr

*Corresponding author

Financial support: This work was supported by a grant (FAIR CT 97-3097) from the European Community.

Keywords: biological activity, cyclic AMP, peptone, radioimmunoassay, radioreceptorassay.

Abstract Full Text


Calcitonin gene related peptide (CGRP) related molecules were purified from sardine hydrolysates prepared using 0.1% alcalase and two hours of hydrolysis. Gel exclusion chromatography and HPLC performed purification of these molecules. The purified molecules were characterised using specific CGRP radioimmunoassays and radioreceptoraasays. From 22 mg of crude extract, we obtained 14 µg of CGRP related molecules, the molecular weight determined by mass spectrophotometry was 6000 daltons. The biological activity of these molecules was analysed using the ability of CGRP to stimulate the adenylate cyclase activity in rat liver membranes. The purified molecules induced an inhibition of the CGRP stimulated adenylate cyclase activity, this effect was specific as no such effect was observed on the glucagon stimulated adenylate cyclase activity measured in the same rat liver membrane preparation. These results suggest that the purified molecules may act as antagonists for peptides that bind to CGRP receptors in rat liver membranes. These new antagonists may be of particular importance in various aspects of CGRP action namely in the control of animal feeding.
Supported by UNESCO / MIRCEN network
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