cDNA cloning of a novel secreted isoform of the human receptor for advanced glycation end products and characterization of cells co-expressing cell-surface scavenger receptors and Swedish mutant amyloid precursor protein
cDNA cloning of a novel secreted isoform of the human receptor for advanced glycation end products and characterization of cells co-expressing cell-surface scavenger receptors and Swedish mutant amyloid precursor protein
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DOI：
10.1016/s0169-328x(99)00174-6
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发表时间：
1999-08-25
期刊：
影响因子：
--
通讯作者：
Huber, G
中科院分区：
文献类型：
--
作者：
Malherbe, P;Richards, JG;Huber, G
已提出晚期糖基化终产物受体（AGEs）作为细胞表面受体，其结合淀粉样β蛋白（A β），从而触发其细胞毒性作用[S. D. Yan，X. Chen，J. Fu，M. Chen，H. Zhu，中国茶青冈A. Roher，T.斯莱特里湖Zhao，M. Nagashima，J. Morser，A. Migheli，P. Nawroth，D. Stem，A.M.施密特，Alzheimer病中的淀粉样蛋白和淀粉样蛋白-P肽神经毒性，Nature 382（1996）685-691.]。用合适的杂交探针对人肺cDNA文库进行筛选。除了细胞表面蛋白外，还发现了一个克隆，它编码一种新的蛋白，称为hRAGEsec，它缺乏跨膜区的19个氨基酸，因此被分泌。与基因组序列的比较表明，分泌型异构体的合成需要选择性剪接。推导的成熟hRAGEsec的蛋白质序列由321个氨基酸组成，预测的分子量为35.66 kDa。用原位杂交组织化学方法分析hRAGEsec在人脑中的表达模式。在海马CA 3区锥体细胞、齿状回颗粒细胞、皮质神经元以及白色物质中的胶质细胞中，该基因的表达最强，而在细胞表面表达最弱。为了研究在生理条件下A β和β-淀粉样蛋白以及另一种清道夫受体SRA之间的相互作用，将它们与人β APP（695）-SFAD在人细胞中共表达，并通过免疫沉淀和酶联免疫吸附测定（ELISA）分析评估条件培养基中A β的水平。A β从hepatocellular细胞的条件培养基中几乎100%的减少和类似于从SRA-细胞中40%的减少暗示hepatocellular可能是与AP相互作用的突出的细胞表面受体。(C)1999 Elsevier Science B. V.保留所有权利。
The receptor for advanced glycation end products (RAGE) has been proposed as a cell surface receptor that binds amyloid-beta protein (A beta), thereby triggering its cytotoxic effects [S.D. Yan, X. Chen, J. Fu, M. Chen, H. Zhu, A. Roher, T. Slattery, L. Zhao, M. Nagashima, J. Morser, A. Migheli, P. Nawroth, D. Stem, A.M. Schmidt, RAGE and amyloid-P peptide neurotoxicity in Alzheimer's disease, Nature 382 (1996) 685-691.]. A cDNA library of human lung was screened for RAGE with an appropriate hybridization probe. In addition to cell surface RAGE, one clone was found which encodes a new version of RAGE, termed hRAGEsec, which lacks the 19 amino acids of the membrane-spanning region and is therefore secreted. Comparison with the genomic sequence revealed that the synthesis of the secreted isoform requires alternative splicing. The deduced protein sequence of the mature hRAGEsec consists of 321 amino acids with a predicted molecular mass of 35.66 kDa. The pattern of expression of hRAGEsec in human brain was analyzed by in situ hybridization histochemistry. The most intense expression of the gene in contrast to cell surface RAGE was detected in hippocampal CA3 pyramidal cells, dentate gyms granule cells, cortical neurons as well, as glial cells in white matter. To investigate the interaction between A beta and RAGE and another scavenger receptor, SRA, under physiological conditions, they were co-expressed with human beta APP(695)-SFAD in a human cell and the level of A beta in the condition medium was assessed by immunoprecipitation and enzyme-linked immunosorbent assay (ELISA) analysis. A nearly 100% reduction of A beta from the conditioned medium of hRAGE cells and similar to 40% reduction from the SRA-cells implied that hRAGE could be a prominent cell surface receptor interacting with AP. (C) 1999 Elsevier Science B.V. All rights reserved.