Harashima Ai, Yamamoto Yasuhiko, Cheng Chunmei, Tsuneyama Koichi, Myint Khin Mar, Takeuchi Akihiko, Yoshimura Kazunobu, Li Hui, Watanabe Takuo, Takasawa Shin, Okamoto Hiroshi, Yonekura Hideto, Yamamoto Hiroshi
Department of Biochemistry and Molecular Vascular Biology, Kanazawa University Graduate School of Medical Science, Kanazawa 920-8640, Japan.
Biochem J. 2006 May 15;396(1):109-15. doi: 10.1042/BJ20051573.
The cell-surface RAGE [receptor for AGE (advanced glycation end-products)] is associated with the development of diabetic vascular complications, neurodegenerative disorders and inflammation. Recently, we isolated a human RAGE splice variant, which can work as a decoy receptor for RAGE ligands, and named it esRAGE (endogenous secretory RAGE). In the present study, we have isolated the murine equivalent of esRAGE from brain polysomal poly(A)+ (polyadenylated) RNA by RT (reverse transcription)-PCR cloning. The mRNA was generated by alternative splicing, and it encoded a 334-amino-acid protein with a signal sequence, but lacking the transmembrane domain. A transfection experiment revealed that the mRNA was actually translated as deduced to yield the secretory protein working as a decoy in AGE-induced NF-kappaB (nuclear factor kappaB) activation. RT-PCR and immunoblotting detected esRAGE mRNA and protein in the brain, lung, kidney and small intestine of wild-type mice, but not of RAGE-null mice. The esRAGE expression was increased in the kidney of diabetic wild-type mice. The present study has thus provided an animal orthologue of esRAGE for clarification of its roles in health and disease.
细胞表面的RAGE [晚期糖基化终末产物(AGE)受体] 与糖尿病血管并发症、神经退行性疾病及炎症的发生发展相关。最近,我们分离出一种人RAGE剪接变体,它可作为RAGE配体的诱饵受体,我们将其命名为esRAGE(内源性分泌型RAGE)。在本研究中,我们通过RT(逆转录)-PCR克隆从脑多核糖体多聚腺苷酸[poly(A)+]RNA中分离出了小鼠的esRAGE同源物。该mRNA通过可变剪接产生,编码一个含有信号序列但缺少跨膜结构域的334个氨基酸的蛋白质。转染实验表明,该mRNA实际按照推导进行翻译,产生在AGE诱导的核因子κB(NF-κB)激活中起诱饵作用的分泌蛋白。RT-PCR和免疫印迹检测到野生型小鼠脑、肺、肾和小肠中有esRAGE mRNA和蛋白,但RAGE基因敲除小鼠中未检测到。糖尿病野生型小鼠肾脏中的esRAGE表达增加。因此,本研究提供了esRAGE的动物同源物,以阐明其在健康和疾病中的作用。