Systematic isolation of peptide signal molecules regulating development in hydra: LWamide and PW families
Systematic isolation of peptide signal molecules regulating development in hydra: LWamide and PW families
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DOI：
10.1073/pnas.94.4.1241
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发表时间：
1997-02-18
影响因子：
11.1
通讯作者：
Sugiyama, T
中科院分区：
文献类型：
--
作者：
Takahashi, T;Muneoka, Y;Sugiyama, T
为了分离新的参与调节水螅发育过程的肽信号分子，开发了一种新的筛选项目。采用高效液相色谱法对从大乳头水螅组织中提取的多肽进行了系统纯化。通过差异显示-PCR检查每个纯化肽的一部分，以确定其影响水螅中基因表达的能力。使用氨基酸序列分析仪和/或质谱仪使用另一个级分来确定暂定结构。基于结果，选择潜在感兴趣的肽用于化学合成，然后使用HPLC确认合成肽与天然肽的同一性。使用这种方法，286肽已被分离，暂定氨基酸序列已被确定为其中95个，和19个合成肽相同的天然的。19种合成肽在多种生物学测试中具有活性。例如，Hym-54刺激了水螅和海葵（Anthopleura fuscoviridis）的成年息肉的肌肉收缩，并诱导了海洋水螅虫（Hydractinia serrata）的幼虫阶段的浮游虫变态为息肉。另一种肽，Hym-33 H，抑制水螅神经细胞的分化，并诱导组织收缩的水螅的浮状体。迄今为止获得的证据表明，水螅含有大量（>350）肽信号分子参与调节刺胞动物的发育或其他过程。这些肽可以被分离，并与本研究中开发的新方法系统地检查其功能。
To isolate new peptide signal molecules involved in regulating developmental processes in hydra, a novel screening project was developed. Peptides extracted from the tissue of Hydra magnipapillata were systematically purified to homogeneity using HPLC. A fraction of each purified peptide was examined by differential display-PCR for its ability to affect gene expression in hydra. Another fraction was used to determine the tentative structure using an amino acid sequence analyzer and/or a mass spectrometer. Based on the results, peptides of potential interest were selected for chemical synthesis, followed by confirmation of the identity of the synthetic with the native peptides using HPLC. Using this approach, 286 peptides have been isolated, tentative amino acid sequences have been determined for 95 of them, and 19 synthetic peptides identical to native ones were produced. The 19 synthetic peptides were active in a variety of biological tests. For example, Hym-54 stimulated muscle contraction in adult polyps of hydra and sea anemone, Anthopleura fuscoviridis, and induced metamorphosis of planula, the larval stage, into polyps in a marine hydrozoan species, Hydractinia serrata. Another peptide, Hym-33H, inhibited nerve cell differentiation in hydra and induced tissue contraction in planula of Hydractinia serrata. The evidence obtained so far suggests that hydra contains a large number (>350) of peptide signal molecules involved in regulating developmental or other processes in cnidaria. These peptides can be isolated and their functions examined systematically with the new approach developed in this study.