De novo sequencing of peptides secreted by the skin glands of the Caucasian Green Frog Rana ridibunda
De novo sequencing of peptides secreted by the skin glands of the Caucasian Green Frog Rana ridibunda
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DOI：
10.1002/rcm.3759
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发表时间：
2008-11-01
影响因子：
2
通讯作者：
Lebedev, Albert T.
中科院分区：
文献类型：
--
作者：
Samgina, Tatiana Yu.;Artemenko, Konstantin A.;Lebedev, Albert T.
已知两栖动物的皮肤腺分泌各种类型的生物活性肽。这些肽的阵列对每种青蛙都是特异性的。本研究对沼泽蛙R.居住在高加索地区科尔希达峡谷的海脊鱼。该研究基于对完整和化学修饰肽的全面高效液相色谱/串联质谱（HPLC/MS/MS）分析。特别地，将氧化程序直接应用于粗皮肤分泌物以打开S-S环，而另外对一个单独的肽进行N-末端乙酰化。通过手动解释电子捕获解离（ECD）和碰撞诱导解离（CID）串联质谱确定序列。在高加索沼泽蛙皮肤分泌物中共鉴定出29种多肽。肽谱用属于短肽蛋白、七叶素和拉纳图林家族的含二硫键的肽、缓激肽和铃蟾肽家族的神经肽表示。两种属于Ranatuerins的已鉴定肽是在欧洲青蛙的皮肤分泌物中发现的该家族的第一种肽。其中10种肽与先前报道的欧洲可食用青蛙的肽一致。另外10个与R中发现的相同。来自莫斯科地区。这一事实证明，所描述的方法是一种有效的分析工具，比较种内和种间的变异。版权所有（c）2008约翰威利父子有限公司。
Amphibian skin glands are known to secrete various types of bioactive peptides. The array of these peptides is specific for every frog species. The present research deals with the identification of peptides isolated from the skin secretion of the Marsh frog R. ridibunda inhabiting the Kolkhida Canyon of the Caucasian region. The research is based on comprehensive high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) analysis of intact and chemically modified peptides. In particular, an oxidation procedure was applied directly to the crude skin secretion to open S-S loops whereas N-terminal acetylation was additionally carried out for one individual peptide. Sequences were determined by manual interpretation of electron capture dissociation (ECD) and collisionally induced dissociation (CID) tandem mass spectra. A total of 29 peptides were identified in the skin secretion of the Caucasian Marsh frog. The peptide profile is represented with disulfide-containing peptides belonging to the brevinin, esculentin and ranatuerin families, neuropeptides of the bradykinin and bombesin families. Two identified peptides belonging to the ranatuerins are the first peptides of this family discovered in the skin secretions of European frogs. Ten of the identified peptides coincide with those reported earlier for the European Edible frog. Another ten are identical to those found in R. ridubunda from the Moscow region. This fact verifies the described method as being art efficient analytical tool to compare intra- and interspecific variabilities. Copyright (c) 2008 John Wiley & Sons, Ltd.