Distribution of CESP-1 protein in the corneal endothelium and other tissues
Distribution of CESP-1 protein in the corneal endothelium and other tissues
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DOI：
10.1167/iovs.05-0602
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发表时间：
2006-04-01
影响因子：
4.4
通讯作者：
Yamagami, S
中科院分区：
文献类型：
--
作者：
Kinouchi, R;Kinouchi, T;Yamagami, S
目的。利用基因签名系统建立了人角膜内皮（CE）的基因表达谱。根据人类基因表达数据库，与其他组织相比，一种新基因 GS3582 在 CE 中转录丰富。该蛋白被命名为角膜内皮特异性蛋白(CESP)-1。评估CESP-1在人和小鼠体内的组织分布和亚细胞定位，探讨其生理功能。方法。克隆了兔和小鼠 CESP-1 cDNA，并产生了多克隆抗人 CESP-1 抗体 (Ab) 和抗小鼠 N 端或 C 端卵巢特异性酸性蛋白 (OSAP)-1 Ab。通过蛋白质印迹和/或免疫组织化学分析研究人和小鼠角膜中的 CESP-1 表达。还通过 Western Not 分析检查了 CESP-1 在人体组织中的分布。为了确定 CESP-1 的亚细胞定位，将培养的人 CE 与抗人 CESP-1 Ab 和抗细胞色素 c 单克隆抗体或抗 GRP78 单克隆抗体共同标记，用于共聚焦显微镜检查。结果。兔和小鼠CESP-1 cDNA序列分别包含编码242和283个氨基酸的开放阅读框。小鼠CESP-1与小鼠OSAP完全一致。 Western blot分析显示CESP-1在人角膜上皮、CE、培养的CE、脑、睾丸和卵巢中表达。根据免疫组织化学分析，小鼠 CESP-1 也在小鼠角膜上皮和 CE 中表达，并且具有抗小鼠 C 端而非 N 端 OSAP 抗体。在培养的人类 CE 中证实了 CESP-1 向线粒体的亚细胞定位。 CESP-1的N端具有线粒体靶向序列，可在蛋白质导入线粒体后进行加工。结论。 CESP-1分布于角膜上皮、CE和培养的人类CE以及大脑、睾丸和卵巢中。 CESP-1 定位于培养的人类 CE 的线粒体中。这些发现可能为一些城市提供有关CESP-1生理功能的信息。
PURPOSE. The gene expression profile of human corneal endothelium (CE) was established with the gene signature system. A novel gene, GS3582, was abundantly transcribed in the CE compared with other tissues according to a human gene expression database. This protein was designated corneal endothelium -specific protein (CESP)-1. The tissue distribution and subcellular localization of CESP-1 was assessed in humans and mice, to investigate its physiological function.METHODS. Rabbit and mouse CESP-1 cDNAs were cloned, and a polyclonal anti-human CESP-1 antibody (Ab) and anti-mouse N- or C-terminal ovary-specific acidic protein (OSAP)-1 Ab were produced. CESP-1 expression was investigated in human and mouse corneas by Western blot and/or immunohistochemical analysis. The distribution of CESP-1 in human tissues was also examined by Western Not analysis. To identify the subcellular localization of CESP-1, cultured human CE was colabeled with anti-human CESP-1 Ab and anti-cytochrome c monoclonal Ab or anti-GRP78 monoclonal Ab for confocal microscopy.RESULTS. The rabbit and mouse CESP-1 cDNA sequences contained an open reading frame coding 242 and 283 amino acids, respectively. Mouse CESP-1 was entirely consistent with mouse OSAP. Western blot analysis showed that CESP-1 was expressed in the human corneal epithelium, CE, cultured CE, brain, testis, and ovary. Mouse CESP-1 was also expressed in mouse corneal epithelium and CE with anti-mouse C- but not N-terminal OSAP Ab according to immunohistochemical analysis. Subcellular localization of CESP-1 to the mitochondria was demonstrated in cultured human CE. The N-terminal of CESP-1, possessing a mitochondrial targeting sequence, may be processed after the protein is imported into the mitochondria.CONCLUSIONS. CESP-1 was distributed in the corneal epithelium, the CE and cultured human CE, as well as the brain, testis, and ovary. CESP-1 was localized in the mitochondria of cultured human CE. These findings may provide some cities about the physiological function of CESP-1.