Molecular cloning of a novel P2 purinoceptor from human erythroleukemia cells
Molecular cloning of a novel P2 purinoceptor from human erythroleukemia cells
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DOI：
10.1074/jbc.271.31.18363
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发表时间：
1996-08-02
影响因子：
4.8
通讯作者：
Kunapuli, SP
中科院分区：
文献类型：
--
作者：
Akbar, GKM;Dasari, VR;Kunapuli, SP
用放射性标记的鸡P_2Y（3）cDNA低严谨度筛选人红白血病细胞cDNA文库，发现一个编码与P_2嘌呤受体同源的G蛋白偶联受体的cDNA克隆，命名为P_2Y（7），该受体具有352个氨基酸，与P_2Y（1）-P_2Y（6）嘌呤受体具有23-30%的氨基酸同源性。P2 Y（7）cDNA在COS-7细胞中瞬时表达：其结合研究显示对ATP具有非常高的亲和力（37 +/- 6 nM），UTP和ADP的浓度更低（类似于1300 nM），以及在8个核苷酸和苏拉明的结合系列研究中的新的亲和力等级顺序，P2 Y（7）受体序列似乎表示与所有其他已知P2 Y嘌呤受体不同的亚家族，只有少数特征性序列基序是共享的，P2 Y（7）受体mRNA在人心脏和骨骼肌中大量存在，在脑和肝中中等程度存在，但在其他测试组织中不存在。P2 Y（7）受体mRNA在大鼠心脏和培养的新生大鼠心肌细胞中也大量存在，P2 Y（7）受体在瞬时表达该受体的COS-7细胞中与磷脂酶C功能性偶联。P2 Y（7）基因显示定位于人14号染色体。因此，我们克隆了一个独特的成员的P2 Y嘌呤受体家族可能发挥作用，在调节心肌收缩。
Screening of a human erythroleukemia cell cDNA library with radiolabeled chicken P2Y(3) cDNA at low stringency revealed a cDNA clone encoding a novel G protein-coupled receptor with homology to P2 purinoceptors, This receptor, designated P2Y(7), has 352 amino acids and shares 23-30% amino acid identity with the P2Y(1)-P2Y(6) purinoceptors. The P2Y(7) cDNA was transiently expressed in COS-7 cells: binding studies thereon showed a very high affinity for ATP (37 +/- 6 nM), much less for UTP and ADP (similar to 1300 nM), and a novel rank order of affinities in the binding series studied of 8 nucleotides and suramin, The P2Y(7) receptor sequence appears to denote a different subfamily from that of all the other known P2Y purinoceptors, with only a few of their characteristic sequence motifs shared, The P2Y(7) receptor mRNA is abundantly present in the human heart and the skeletal muscle, moderately in the brain and liver, but not in the other tissues tested, The P2Y(7) receptor mRNA was also abundantly present in the rat heart and cultured neonatal rat cardiomyocytes, The P2Y(7) receptor is functionally coupled to phospholipase C in COS-7 cells transiently expressing this receptor, The P2Y(7) gene was shown to be localized to human chromosome 14. We have thus cloned a unique member of the P2Y purinoceptor family which probably plays a role in the regulation of cardiac muscle contraction.