Biochemistry Department, Center for Pharmaceuticals Research and Development, Ave. 26, No. 1605, CP 10 400 Habana, Cuba.
Fish Shellfish Immunol. 2012 Oct;33(4):872-9. doi: 10.1016/j.fsi.2012.07.013. Epub 2012 Aug 5.
Naturally occurring antimicrobial peptides take place in the first line of host defense against pathogen as part of the humoral innate immune response. β-defensins are among the most abundant antimicrobial peptides in mammals, and thought to be solely found in vertebrates until a recent report describing the cloning and sequencing of defensin like peptides in the spiny lobster Panulirus japonicus. In the current study, we cloned and sequenced two genes from the hemocytes of the spiny lobster Panulirus argus encoding for two isoforms of defensin-like peptides, thus confirming the presence of this protein in the Panulirus genus. The 44 amino acids mature peptides showed the conservation of cysteine pattern characterizing the β-defensins, as well as known amino acids residues critical to exert their antimicrobial activity. They are also amphipathics, hydrophobics, and display an overall positive charge (+1) located at the C-terminus. The tertiary structure obtained by homology modeling indicated that likely conformations of lobster peptides are highly similar to β-defensins from vertebrates. The phylogenetic study carried out by probabilistic methods confirmed the relation with ancestral β-defensin from vertebrates. The finding of a putative defensin-like peptide in the expressed sequence tag (EST) of the lobster Homarus americanus with high homology with those of P. argus described in this study, would indicate the presence of this peptides in Palinuridae family. Taking into account all similarities between these peptides with β-defensins from vertebrates, it is conceivable to further support the finding of a new family of β-defensins in invertebrate.
天然存在的抗菌肽作为体液先天免疫反应的一部分,发生在宿主防御病原体的第一线。β-防御素是哺乳动物中最丰富的抗菌肽之一,直到最近的一份报告描述了在刺龙虾 Panulirus japonicus 中克隆和测序防御素样肽,人们才认为它们仅存在于脊椎动物中。在本研究中,我们从刺龙虾 Panulirus argus 的血细胞中克隆和测序了两个基因,编码两种防御素样肽的同工型,从而证实了这种蛋白质在 Panulirus 属中的存在。成熟的 44 个氨基酸肽显示出保守的半胱氨酸模式,特征是β-防御素,以及发挥其抗菌活性的关键氨基酸残基。它们也是两性的、疏水性的,并在 C 末端显示出整体正电荷 (+1)。通过同源建模获得的三级结构表明,龙虾肽的可能构象与脊椎动物的β-防御素高度相似。通过概率方法进行的系统发育研究证实了与脊椎动物祖先β-防御素的关系。在表达序列标签 (EST) 中发现了一种假定的防御素样肽在龙虾 Homarus americanus 中,与本研究中描述的 P. argus 具有高度同源性,这表明这种肽在 Palinuridae 科中存在。考虑到这些肽与脊椎动物β-防御素之间的所有相似性,可以进一步支持在无脊椎动物中发现新的β-防御素家族的发现。