Purification and characterization of avian beta-defensin 11, an antimicrobial peptide of the hen egg.
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.
Hervé-Grépinet
Virginie
V
INRA, UR83 Recherches Avicoles, F-37380 Nouzilly, France.
Réhault-Godbert
Sophie
S
Labas
Valérie
V
Magallon
Thierry
T
Derache
Chrystelle
C
Lavergne
Marion
M
Gautron
Joël
J
Lalmanach
Anne-Christine
AC
Nys
Yves
Y
D000818:Animals; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D015153:Blotting, Western; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004591:Electrophoresis, Polyacrylamide Gel; D008089:Listeria monocytogenes / Q000187:drug effects; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014817:Vitelline Membrane / Q000737:chemistry; D023083:beta-Defensins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D010455:Peptides; D023083:beta-Defensins