Cloning and characterization of human protease-activated receptor 4
Cloning and characterization of human protease-activated receptor 4
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DOI：
10.1073/pnas.95.12.6642
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发表时间：
1998-06-09
影响因子：
11.1
通讯作者：
Foster, DC
中科院分区：
文献类型：
--
作者：
Xu, WF;Andersen, H;Foster, DC
蛋白酶激活受体1-3 （PAR1、PAR2和PAR3）是一个独特的G蛋白偶联受体家族的成员。在表达的序列标签数据库中鉴定了该家族第4个成员（PAR4）的部分cDNA序列，并从淋巴瘤Daudi细胞cDNA文库中分离出全长cDNA克隆。ORF编码7个跨膜结构域蛋白，包含385个氨基酸，其中33%的氨基酸序列与PAR1、PAR2和PAR3相同。在胞外氨基端确定了一个假定的蛋白酶裂解位点（Arg-47/Gly-48）。短暂转染PAR4的COS细胞在凝血酶或胰蛋白酶处理下均可形成细胞内肌醇三磷酸，其中Arg-47被Ala取代的PAR4突变体对凝血酶或胰蛋白酶没有反应。经凝血酶激活的COS细胞转染野生型或突变型PARA Northern blot进行蛋白水解后，来自PAR4氨基端新暴露的拴链配体的六肽（GYPGQV）表明，PAR4 mRNA在许多人体组织中表达，在肺、胰腺、甲状腺、睾丸和小肠中均有高水平表达。利用荧光原位杂交技术，将PAR4基因定位到19p12染色体上。
Protease-activated receptors 1-3 (PAR1, PAR2, and PAR3) are members of a unique G protein-coupled receptor family. They are characterized by a tethered peptide ligand at the extracellular amino terminus that is generated by minor proteolysis, A partial cDNA sequence of a fourth member of this family (PAR4) was identified in an expressed sequence tag database, and the full-length cDNA clone has been isolated from a lymphoma Daudi cell cDNA library. The ORF codes for a seven transmembrane domain protein of 385 amino acids with 33% amino acid sequence identity with PAR1, PAR2, and PAR3. A putative protease cleavage site (Arg-47/Gly-48) was identified within the extracellular amino terminus. COS cells transiently transfected with PAR4 resulted in the formation of intracellular inositol triphosphate when treated with either thrombin or trypsin, A PAR4 mutant in which the Arg-47 was replaced with Ala did not respond to thrombin or trypsin, A hexapeptide (GYPGQV) representing the newly exposed tethered ligand from the amino terminus of PAR4 after proteolysis by thrombin activated COS cells transfected with either wild-type or the mutant PARA Northern blot showed that PAR4 mRNA was expressed in a number of human tissues, with high levels being present in lung, pancreas, thyroid, testis, and small intestine. By fluorescence in situ hybridization, the human PAR4 gene was mapped to chromosome 19p12.