Dugimont T, Guissi-Kadri S, Curgy J J
Laboratory of Invertebrate Endocrinology, CNRS UA 148, University of Lille-Flandres-Artois, Villeneuve d'Ascq, France.
Int J Pept Protein Res. 1992 Apr;39(4):300-7. doi: 10.1111/j.1399-3011.1992.tb01589.x.
Total mRNA were extracted from brain of Nereis diversicolor (Annelida, Polychaeta) and were translated in vitro or in ovo. The newly synthesized polypeptides were analyzed through electrophoresis of immunoprecipitated products or the Western blotting technique using polyclonal antibodies raised against mammalian dynorphin 1-17 and mammalian alpha-neo-endorphin. Among the products translated in vitro, only one class of polypeptide of 70 kDa was recognized by anti-dynorphin 1-17 antibodies. Furthermore, some in ovo translated products as well as proteins extracted from brain of worms showed identical immunoreactivity. These polypeptides, 60-70 kDa, reacted with anti-dynorphin 1-17 and anti alpha-neo-endorphin antibodies. These results suggest the existence of epitopes common to in ovo and in vitro translated products, to polypeptides extracted from the brain and to some mammalian opioid peptides of the prodynorphin family. We postulate the presence, in the brain of N. diversicolor, of precursors of peptides related to mammalian dynorphin 1-17 and alpha-neo-endorphin. Data reported in this investigation do not allow us to propose or even postulate the presence, in the brain of the worm, of one precursor molecule common to polypeptides related to mammalian dynorphin 1-17 and alpha-neo-endorphin. Furthermore, the Nereis precursor molecules exhibit a clear-cut difference in molecular mass with the mammalian prodynorphin: 70 kDa versus 30 kDa.
从杂色沙蚕(环节动物门,多毛纲)的脑中提取总mRNA,并在体外或卵内进行翻译。使用针对哺乳动物强啡肽1-17和哺乳动物α-新内啡肽产生的多克隆抗体,通过免疫沉淀产物的电泳或蛋白质印迹技术对新合成的多肽进行分析。在体外翻译的产物中,只有一类70 kDa的多肽能被抗强啡肽1-17抗体识别。此外,一些卵内翻译产物以及从蠕虫脑中提取的蛋白质显示出相同的免疫反应性。这些60 - 70 kDa的多肽与抗强啡肽1-17和抗α-新内啡肽抗体发生反应。这些结果表明,卵内和体外翻译产物、从脑中提取的多肽以及前强啡肽家族的一些哺乳动物阿片肽存在共同的表位。我们推测在杂色沙蚕的脑中存在与哺乳动物强啡肽1-17和α-新内啡肽相关的肽前体。本研究报告的数据不允许我们提出甚至推测在蠕虫脑中存在与哺乳动物强啡肽1-17和α-新内啡肽相关的多肽共同的一种前体分子。此外,沙蚕前体分子与哺乳动物前强啡肽在分子量上有明显差异:分别为70 kDa和30 kDa。