Purification and Characterization of Avian β-Defensin 11, an Antimicrobial Peptide of the Hen Egg
Purification and Characterization of Avian β-Defensin 11, an Antimicrobial Peptide of the Hen Egg
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DOI：
10.1128/aac.00204-10
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发表时间：
2010-10-01
影响因子：
4.9
通讯作者：
Nys, Yves
中科院分区：
文献类型：
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作者：
Herve-Grepinet, Virginie;Rehault-Godbert, Sophie;Nys, Yves
天然抗菌肽存在于鸡蛋的不同隔室（蛋壳、蛋清和卵黄膜）中，有望参与胚胎发育过程中的保护，并有助于生产无病原体的鸡蛋。在本研究中，我们使用母鸡（Gallus Gallus）鸡蛋的卵黄膜作为禽β -防御素11 （AvBD11）的来源。建立了亲和层析和反相层析的纯化方案。对纯化的AvBD11进行质谱分析，确定其一级序列和结构。在[M + H]（+）上获得了9271.56 Da的单同位素分子，并确定了其N端和c端序列。我们还检查了翻译后修饰，并确定了6个内部二硫键的存在。AvBD11对革兰氏阳性菌和革兰氏阴性菌均有抗菌活性。
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 beta-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.