MOLECULAR-CLONING AND EXPRESSION OF MURINE AND BOVINE ENDOTHELIAL-CELL PROTEIN C/ACTIVATED PROTEIN-C RECEPTOR (EPCR) - THE STRUCTURAL AND FUNCTIONAL CONSERVATION IN HUMAN, BOVINE, AND MURINE EPCR
MOLECULAR-CLONING AND EXPRESSION OF MURINE AND BOVINE ENDOTHELIAL-CELL PROTEIN C/ACTIVATED PROTEIN-C RECEPTOR (EPCR) - THE STRUCTURAL AND FUNCTIONAL CONSERVATION IN HUMAN, BOVINE, AND MURINE EPCR
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DOI：
10.1074/jbc.270.10.5571
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发表时间：
1995-03-10
影响因子：
4.8
通讯作者：
ESMON, CT
中科院分区：
文献类型：
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作者：
FUKUDOME, K;ESMON, CT
最近，我们鉴定并克隆了一种人内皮细胞蛋白C/活化蛋白C受体（EPCR），预测EPCR是一种1型跨膜糖蛋白，是CD 1/主要组织相容性复合物超家族的新成员，与CD 1d具有28%的同源性。甚至更大的同源性（62%的同一性），检测到与鼠蛋白质，CCD 41，这是以前的特点是作为一个中心体相关的，细胞周期依赖性蛋白。这提出了CCD 41是EPCR的鼠同源物的可能性。为了解决这种可能性，以更好地了解结构-功能关系，并促进EPCR功能的生理实验，我们克隆和测序小鼠和牛EPCR从内皮细胞cDNA文库。小鼠EPCR和CCD 41的核苷酸序列表现出五个差异，对应于蛋白质编码区中的一个碱基变化，三个单碱基插入和一个碱基缺失。结果表明，EPCR和CCD 41的预测结构在其氨基和羧基末端不同，但在编码序列的中心部分相同。基于鼠、牛和人EPCR序列的比较以及检测到鼠EPCR和CCD 41之间差异的区域，我们认为CCD 41可能与鼠EPCR相同，并且报告的序列差异可能是测序凝胶上压缩的结果。与人EPCR相比，鼠EPCR和牛EPCR的同源性分别为69%和73%，3个物种之间的同源性为57%。与人EPCR一样，在所测试的细胞系中，鼠EPCR信息仅限于内皮。克隆鼠和牛EPCR同源物将有助于在体内和体外研究EPCR在蛋白C途径中的作用。
Recently, we identified and cloned a human endothelial cell protein C/activated protein C receptor (EPCR), EPCR was predicted to be a type 1 transmembrane glycoprotein and a novel member of the CD1/major histocompatibility complex superfamily with 28% identity with CD1d. Even greater homology (62% identity) was detected with the murine protein, CCD41, which was previously characterized as a centrosome associated, cell cycle-dependent protein. This raised the possibility that CCD41 was the murine homologue of EPCR. To address this possibility, to better understand structure-function relationships, and to facilitate physiological experiments on EPCR function, we cloned and sequenced murine and bovine EPCR from endothelial cell cDNA libraries. The nucleotide sequence of murine EPCR and CCD41 exhibited five differences corresponding to one base change, three single-base insertions, and one base deletion in the protein coding region. As a result, the predicted structures of EPCR and CCD41 differed in their amino and carboxyl termini but were identical in the central portion of the coding sequence. Based on comparison of the murine, bovine, and human EPCR sequences and the regions where discrepancies between murine EPCR and CCD41 were detected, we believe that CCD41 is probably identical to murine EPCR and that the reported sequence differences are likely the result of compression on the sequencing gel. Compared with human EPCR, the murine and bovine sequences were 69 and 73% identical, respectively, and 57% of the residues were identical between all three species, Both bovine and murine EPCR could bind human activated protein C when the cDNA clones were transfected into 293T cells. Like human EPCR, of the cell Lines tested, the murine EPCR message was restricted to endothelium, Cloning of the murine and bovine homologue of EPCR will facilitate in vivo and in vitro studies of the role of EPCR in the protein C pathway.