van Dijk Albert, Veldhuizen Edwin J A, van Asten Alphons J A M, Haagsman Henk P
Department of Public Health and Food Safety, Graduate School of Animal Health, Faculty of Veterinary Medicine, Utrecht University, 3508 TD Utrecht, The Netherlands.
Vet Immunol Immunopathol. 2005 Jul 15;106(3-4):321-7. doi: 10.1016/j.vetimm.2005.03.003. Epub 2005 Apr 7.
Cathelicidins, antimicrobial peptides with broad spectrum activity, have been almost exclusively found in mammals. Here, we report the cloning of a novel avian cathelicidin, chicken myeloid antimicrobial peptide 27 (CMAP27) from chicken bone marrow cells. A combined expressed sequence tag (EST) and genomic based search revealed a cathelicidin-like gene located at the terminus of chromosome 2. Reverse transcriptase-polymerase chain reaction (RT-PCR) and 5'RACE techniques resulted in a 154 amino acid prepropeptide, homologous to chicken cathelicidin 1 (51%) and most similar to alpha-helical myeloid antibacterial peptides (MAPs; 29-33%). A putative elastase cleavage site (LVQRG/RF) suggests the production of a 27 amino acid antimicrobial peptide, predicted to adopt an alpha-helical configuration followed by a hydrophobic tail. Comparative analyses between antimicrobial peptide domains showed marked similarity between CMAP27 and MAP members of the bovidae family, but not with the alpha-helical chicken cathelicidin 1. Strongest expression of CMAP27 mRNA was found in myeloid/lymphoid tissues, testis and uropygial gland. In accordance with the phylogenetic tree analysis, these findings support the theory of a common ancestral cathelicidin gene and suggest an important role for cathelicidins in chicken innate host defense.
防御素是一类具有广谱活性的抗菌肽,几乎仅在哺乳动物中发现。在此,我们报告了一种新型禽类防御素——鸡骨髓抗菌肽27(CMAP27)的克隆,该抗菌肽来自鸡骨髓细胞。通过结合表达序列标签(EST)和基于基因组的搜索,发现一个类防御素基因位于2号染色体末端。逆转录聚合酶链反应(RT-PCR)和5'RACE技术得到了一个154个氨基酸的前原肽,它与鸡防御素1同源(51%),与α-螺旋骨髓抗菌肽(MAPs;29-33%)最为相似。一个假定的弹性蛋白酶切割位点(LVQRG/RF)表明会产生一种27个氨基酸的抗菌肽,预计其采用α-螺旋结构,后面跟着一个疏水尾巴。抗菌肽结构域之间的比较分析表明,CMAP27与牛科动物的MAP成员有显著相似性,但与α-螺旋鸡防御素1没有相似性。CMAP27 mRNA在骨髓/淋巴组织、睾丸和尾脂腺中表达最强。根据系统发育树分析,这些发现支持了防御素基因有共同祖先的理论,并表明防御素在鸡先天宿主防御中起重要作用。