Zaballos Angel, Villares Ricardo, Albar Juan Pablo, Martínez-A Carlos, Márquez Gabriel
Departamento de Inmunología y Oncología, Centro Nacional de Biotecnología/Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Cantoblanco, E-28049 Madrid, Spain.
J Biol Chem. 2004 Mar 26;279(13):12421-6. doi: 10.1074/jbc.M307697200. Epub 2004 Jan 12.
Defensins are important elements in innate immunity that can also trigger adaptive immune responses. The defensins form a family of small cationic antimicrobial peptides with six characteristic cysteine residues, whose pairing pattern in forming three intramolecular disulfide bonds defines the alpha- and beta-defensin subfamilies. In a search for new beta-defensin genes, we performed computational analysis using the Celera mouse genome data base and found exons encoding 23 different beta-defensins, including the eight previously characterized members of this family. Among the new beta-defensins, nine of them form two groups of phylogenetically related sequences that were characterized in greater detail. Northern blot, reverse transcription PCR, and in situ hybridization analysis showed that expression of these genes is restricted to the epididymis, with a specific regional expression pattern. One of the new beta-defensins (Defb38) was chemically synthesized; in in vitro assays on Gram-positive and -negative bacterial strains, Defb38 showed the characteristic salt-dependent antimicrobial activity of beta-defensins. The results demonstrate the existence of a relatively large number of beta-defensins with specific expression in distinct regions of the murine epididymis and suggest complex roles for these proteins in host defense and other physiological processes of the male reproductive tract.
防御素是固有免疫中的重要成分,也可触发适应性免疫反应。防御素构成了一类具有六个特征性半胱氨酸残基的小阳离子抗菌肽家族,其在形成三个分子内二硫键时的配对模式定义了α-防御素和β-防御素亚家族。在寻找新的β-防御素基因的过程中,我们利用赛雷拉小鼠基因组数据库进行了计算分析,发现了编码23种不同β-防御素的外显子,包括该家族先前已鉴定的八个成员。在新的β-防御素中,其中九个形成了两组系统发育相关的序列,并对其进行了更详细的表征。Northern印迹、逆转录PCR和原位杂交分析表明,这些基因的表达仅限于附睾,具有特定的区域表达模式。其中一种新的β-防御素(Defb38)通过化学合成;在对革兰氏阳性和阴性细菌菌株的体外试验中,Defb38表现出β-防御素特有的盐依赖性抗菌活性。结果表明,在小鼠附睾的不同区域存在相对大量具有特异性表达的β-防御素,并提示这些蛋白质在宿主防御和雄性生殖道的其他生理过程中发挥复杂作用。