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Antimicrobial Agents and Chemotherapy, November 2009, p. 4647-4655, Vol. 53, No. 11
0066-4804/09/$08.00+0     doi:10.1128/AAC.00301-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Primary Structure and Antibacterial Activity of Chicken Bone Marrow-Derived β-Defensins{triangledown}

Chrystelle Derache,1 Valérie Labas,2 Vincent Aucagne,3 Hervé Meudal,3 Céline Landon,3 Agnès F. Delmas,3 Thierry Magallon,2 and Anne-Christine Lalmanach1*

INRA, UR 1282 Infectiologie Animale et Santé Publique,1 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,2 Centre de Biophysique Moléculaire, CNRS UPR4301, affiliated to University of Orléans, rue Charles Sadron, 45071 Orléans Cedex 2, France3

Received 5 March 2009/ Returned for modification 2 June 2009/ Accepted 28 August 2009

Three biologically active β-defensins were purified by chromatography from chicken bone marrow extract: avian β-defensin 1 (AvBD1), AvBD2, and the newly isolated β-defensin AvBD7. Mass spectrometry analyses showed that bone marrow-derived AvBD1, -2, and -7 peptides were present as mature peptides and revealed posttranslational modifications for AvBD1 and AvBD7 in comparison to their in silico-predicted amino acid sequences. Tandem mass spectrometry analysis using the nanoelectrospray-quadrupole time of flight method showed N-terminal glutaminyl cyclization of mature AvBD7 and C-terminal amidation of mature AvBD1 peptide, while posttranslational modifications were absent in bone marrow-derived mature AvBD2 peptide. Furthermore, mass spectrometry analysis performed on intact cells confirmed the presence of these three peptides in mature heterophils. In addition, the antibacterial activities of the three β-defensins against a large panel of gram-positive and -negative bacteria were assessed. While the three defensins displayed similar antibacterial spectra of activity against gram-positive strains, AvBD1 and AvBD7 exhibited the strongest activity against gram-negative strains in comparison to AvBD2.

* Corresponding author. Mailing address: INRA, UR 1282 Infectiologie Animale et Santé Publique, F-37380 Nouzilly, France. Phone: 332 47 42 77 00. Fax: 332 47 42 77 74. E-mail: Anne-Christine.Lalmanach{at}tours.inra.fr

{triangledown} Published ahead of print on 8 September 2009.

Antimicrobial Agents and Chemotherapy, November 2009, p. 4647-4655, Vol. 53, No. 11
0066-4804/09/$08.00+0     doi:10.1128/AAC.00301-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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