Mádi András, Mikkat Stefan, Ringel Bruno, Ulbrich Markus, Thiesen Hans-Jürgen, Glocker Michael O
Proteome Center Rostock, Medical Faculty, University of Rostock, Joachim-Jungius-Strasse 9, D-18059 Rostock, Germany.
Proteomics. 2003 Aug;3(8):1526-34. doi: 10.1002/pmic.200300490.
We investigated the effect of cultivation temperatures on the protein expression levels in the fourth larval stage of the postembryonic development of wild-type Caenorhabditis elegans by mass spectrometric proteome analysis. From the 64 protein spots that were investigated, 5 spots were found reproducibly differently expressed when proteome maps derived from animals kept at 15 degrees C and at 25 degrees C, respectively, were compared. Spots of heat shock proteins HSP 70 (CE18679 or CE09682) and HSP 16 (CE14249) were present only in gels from protein extracts when worms were grown at 15 degrees C. Spots of two metabolic enzymes, the isocitrate dehydrogenase (CE10345) and the aspartic proteinase (CE21681) were detected only in cultures grown at the lower temperature as well. A protein with still unknown function (CE05036) was present only in gels from worm samples grown at 25 degrees C. We show for the first time by proteome analyses that cultivation of worms at the lowest temperature of the known physiological range (15 degrees C) already triggers a (weak) stress response in wild-type animals. This work led to the identification of "internal control proteins" in the wild-type strain for further characterization of temperature-sensitive strains using a proteomics approach.
我们通过质谱蛋白质组分析，研究了培养温度对野生型秀丽隐杆线虫胚胎后期发育第四幼虫阶段蛋白质表达水平的影响。在研究的64个蛋白质斑点中，当比较分别在15℃和25℃饲养的动物的蛋白质组图谱时，发现有5个斑点的表达存在可重复的差异。热休克蛋白HSP 70（CE18679或CE09682）和HSP 16（CE14249）的斑点仅出现在15℃培养线虫时提取的蛋白质凝胶中。两种代谢酶，异柠檬酸脱氢酶（CE10345）和天冬氨酸蛋白酶（CE21681）的斑点也仅在较低温度培养的线虫中检测到。一种功能仍未知的蛋白质（CE05036）仅出现在25℃培养线虫的样品凝胶中。我们首次通过蛋白质组分析表明，在已知生理范围的最低温度（15℃）培养线虫，已在野生型动物中引发了（微弱的）应激反应。这项工作促成了野生型菌株中“内部对照蛋白”的鉴定，以便使用蛋白质组学方法进一步表征温度敏感菌株。
