Ostendorp Thorsten, Weibel Mirjam, Leclerc Estelle, Kleinert Peter, Kroneck Peter M H, Heizmann Claus W, Fritz Günter
Fachbereich Biologie, Mathematisch-Naturwissenschaftliche Sektion, Universität Konstanz, Germany.
Biochem Biophys Res Commun. 2006 Aug 18;347(1):4-11. doi: 10.1016/j.bbrc.2006.04.077. Epub 2006 Jun 21.
RAGE is a multi-ligand receptor involved in various human diseases including diabetes, cancer or Alzheimer's disease. Engagement of RAGE by its ligands triggers activation of key cellular signalling pathways such as the MAP kinase and NF-kappaB pathways. Whereas the main isoform of RAGE is a transmembrane receptor with both extra- and intracellular domains, a secreted soluble isoform (sRAGE), corresponding to the extracellular part only, has the ability to block RAGE signalling and suppress cellular activation. Administration of sRAGE to animal models of cancer or multiple sclerosis blocked successfully tumour growth and the course of the autoimmune disease. These findings demonstrate that sRAGE may have a potential as therapeutic. We present here a fast and simple purification protocol of sRAGE from the yeast Pichia pastoris. The identity of the protein was confirmed by mass spectrometry and Western blot. The protein was N-glycosylated and 95-98% pure as judged by SDS-PAGE.
RAGE是一种多配体受体，参与包括糖尿病、癌症或阿尔茨海默病在内的多种人类疾病。其配体与RAGE结合会触发关键细胞信号通路的激活，如丝裂原活化蛋白激酶（MAP激酶）和核因子κB通路。RAGE的主要异构体是一种具有胞外和胞内结构域的跨膜受体，而一种仅对应于胞外部分的分泌型可溶性异构体（sRAGE）具有阻断RAGE信号传导并抑制细胞活化的能力。将sRAGE给予癌症或多发性硬化症动物模型成功阻断了肿瘤生长和自身免疫性疾病的进程。这些发现表明sRAGE可能具有治疗潜力。我们在此展示一种从巴斯德毕赤酵母中快速简单纯化sRAGE的方案。通过质谱和蛋白质免疫印迹法确认了该蛋白质的身份。经十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）判断，该蛋白质进行了N - 糖基化修饰且纯度为95 - 98%。
