Purification and Characterization of Avian β-Defensin 11, an Antimicrobial Peptide of the Hen Egg

Author:

Hervé-Grépinet Virginie1,Réhault-Godbert Sophie1,Labas Valérie2,Magallon Thierry2,Derache Chrystelle3,Lavergne Marion1,Gautron Joël1,Lalmanach Anne-Christine3,Nys Yves1

Affiliation:

1. INRA, UR83 Recherches Avicoles, F-37380 Nouzilly, France

2. UMR INRA 85, CNRS 6175, Université François Rabelais, Physiologie de la Reproduction et des Comportements, Plate-forme de Protéomique Analytique et Fonctionnelle, F-37380 Nouzilly, France

3. INRA, UR 1282 Infectiologie Animale et Santé Publique, F-37380 Nouzilly, France

Abstract

ABSTRACT Natural antimicrobial peptides are present in different compartments (eggshell, egg white, and vitelline membranes) of the hen egg and are expected to be involved in the protection of the embryo during its development and to contribute to the production of pathogen-free eggs. In the present study, we used vitelline membranes from hen ( Gallus gallus ) eggs as a source of avian β-defensin 11 (AvBD11). A purification scheme using affinity chromatography and reverse-phase chromatography was developed. Purified AvBD11 was analyzed by a combination of mass spectrometry approaches to characterize its primary sequence and structure. A monoisotopic molecular species at [M + H] + of 9,271.56 Da was obtained, and its N- and C-terminal sequences were determined. We also examined posttranslational modifications and identified the presence of 6 internal disulfide bonds. AvBD11 was found to exhibit antimicrobial activity toward both Gram-positive and Gram-negative bacteria.

Publisher

American Society for Microbiology

Subject

Infectious Diseases,Pharmacology (medical),Pharmacology

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