Zhou Mei, Chen Tianbao, Walker Brian, Shaw Chris
School of Pharmacy, Queen's University, 97 Lisburn Road, Belfast BT9 7BL, Northern Ireland, UK.
Peptides. 2006 Sep;27(9):2118-23. doi: 10.1016/j.peptides.2006.04.007. Epub 2006 May 19.
Odorous frogs of the sub-genus Odorrana are of oriental distribution, and are so called due to the foul smell of their defensive skin secretions released from specialized skin glands following stress or predator attack. Here we report the application of a "shotgun" skin secretion cDNA library cloning technique which can rapidly expedite identification of secretion bioactive peptides. From a library constructed from the skin secretion of the Large Chinese Odorous frog, Rana (Odorrana) livida, we have identified four novel peptides whose primary structures were deduced initially from cloned precursors. Subsequently, mature peptides were located in and structurally characterized from reverse phase HPLC fractions of skin secretion. Named lividins 1-4, these were found to be structural homologs of known antimicrobial peptide families from Rana frogs. Rapid identification of novel peptides can thus be rapidly achieved using this non-invasive, non-destructive technology and the extensive similarities revealed between antimicrobial peptide precursor organization and nucleic acid sequences would lend support to the hypothesis that they have a common ancestral origin.
臭蛙亚属（Odorrana）的臭蛙分布于东方，因其在受到压力或被捕食者攻击时，会从特化的皮肤腺体释放出具有恶臭的防御性皮肤分泌物而得名。在此，我们报告了一种“鸟枪法”皮肤分泌物cDNA文库克隆技术的应用，该技术可快速加快分泌生物活性肽的鉴定。从由大绿臭蛙（Odorrana livida）的皮肤分泌物构建的文库中，我们鉴定出了四种新型肽，其一级结构最初是从克隆的前体中推导出来的。随后，在皮肤分泌物的反相高效液相色谱馏分中定位并对成熟肽进行了结构表征。这些肽被命名为lividins 1 - 4，发现它们是来自蛙属青蛙的已知抗菌肽家族的结构同源物。因此，使用这种非侵入性、非破坏性技术可以快速鉴定新型肽，并且抗菌肽前体组织与核酸序列之间显示出的广泛相似性将支持它们具有共同祖先起源这一假说。
