Comparison of distinct protein isoforms of the receptor for advanced glycation end-products expressed in murine tissues and cell lines
Comparison of distinct protein isoforms of the receptor for advanced glycation end-products expressed in murine tissues and cell lines
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DOI：
10.1007/s00441-009-0791-0
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发表时间：
2009-07-01
影响因子：
3.6
通讯作者：
Delude, Russell L.
中科院分区：
文献类型：
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作者：
Gefter, Julia V.;Shaufl, Angel L.;Delude, Russell L.
晚期糖基化终产物受体（RAGE）被认为以各种蛋白质异构体的形式普遍表达。我们的目的是使用Northern blotting、免疫blotting和对n -聚糖酶消化的敏感性来调查细胞系和小鼠组织中表达的RAGE亚型，以获得更全面的RAGE表达体视图。肺RAGE mRNA （1.4 kb）小于细胞系和组织RAGE mRNA （6 kb-10 kb）。三种识别三种不同RAGE表位的抗RAGE抗体用于蛋白质研究（N-16， H-300和α ES）。肺表达三种主要的蛋白亚型，表观分子质量分别为45.1、52.6和57.4 kDa (N-16/H-300)，以及四种主要的蛋白亚型，分别为25.0、46.9、52.5和54.2 kDa （alpha ES）。这些亚型仅在肺中表达。心脏、回肠和肾脏表达44.0 kda异构体（N-16），而主动脉和胰腺表达53.3 kda异构体（α ES）。这些异构体在RAGE（-/-）小鼠的组织提取物中均不存在。细胞系表达70.0-kDa亚型，一部分细胞系表达30.0-kDa亚型（α ES）。Lung RAGE似乎含有两种n -链聚糖。组织和细胞系RAGE亚型对PNGase F消化完全不敏感。因此，在组织和细胞系中可以检测到许多RAGE蛋白同工型。典型跨膜和可溶性RAGE似乎仅在肺中表达（N-16/H-300）。非肺组织和细胞系，无论来源组织如何，都表达不同的RAGE蛋白异构体，包含n端N-16表位或由交替外显子9编码的aES RAGE表位，但缺乏H-300表位。
The receptor for advanced glycation end-products (RAGE) is thought to be expressed ubiquitously as various protein isoforms. Our objective was to use Northern blotting, immunoblotting, and sensitivity to N-glycanase digestion to survey RAGE isoforms expressed in cell lines and mouse tissues in order to obtain a more comprehensive view of the RAGE expressome. Pulmonary RAGE mRNA (1.4 kb) was smaller than cell-line and tissue RAGE mRNA (6 kb-10 kb). Three anti-RAGE antibodies that recognized three distinct RAGE epitopes were used for protein studies (N-16, H-300, and alpha ES). Lung expressed three predominant protein isoforms with apparent molecular masses of 45.1, 52.6, and 57.4 kDa (N-16/H-300) and four isoforms at 25.0, 46.9, 52.5, and 54.2 kDa (alpha ES). These isoforms were expressed exclusively in lung. Heart, ileum, and kidney expressed a 44.0-kDa isoform (N-16), whereas aorta and pancreas expressed a 53.3-kDa isoform (alpha ES). Each of these isoforms were absent in tissue extracts prepared from RAGE(-/-) mice. Cell lines expressed a 70.0-kDa isoform, and a subset expressed a 30.0-kDa isoform (alpha ES). Lung RAGE appeared to contain two N-linked glycans. Tissue and cell-line RAGE isoforms were completely insensitive to PNGase F digestion. Thus, numerous RAGE protein isoforms are detectable in tissues and cell lines. Canonical transmembrane and soluble RAGE appear to be expressed solely in lung (N-16/H-300). Non-pulmonary tissues and cell lines, regardless of the source tissue, both express distinct RAGE protein isoforms containing the N-terminal N-16 epitope or the aES RAGE epitope encoded by alternate exon 9, but lacking the H-300 epitope.