Molecular cloning and characterization of a distinct human phosphodiesterase gene family: PDE11A
Molecular cloning and characterization of a distinct human phosphodiesterase gene family: PDE11A
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DOI：
10.1073/pnas.050585197
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发表时间：
2000-03-28
影响因子：
11.1
通讯作者：
Phillips, SC
中科院分区：
文献类型：
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作者：
Fawcett, L;Baxendale, R;Phillips, SC
我们在这里报道了人类PDE11A1的克隆、表达和特征，PDE11A1是一个独特的环核苷酸磷酸二酯酶（PDE）家族的成员。PDE11A与其他所有pde的催化结构域的氨基酸同源性小于或等于50%，与PDE5最相似，具有独特的生化特性。分离的人PDE11A1 cDNA包含一个完整的开放阅读框，编码一个490个氨基酸的酶，预测分子质量为55,786 Da。在N端，PDE11A1具有与其他信号分子（包括PDE2、PDE5、PDE6和PDE10）同源的单个CAF结构域，构成cGMP或其他小配体的潜在变构结合位点。组织分布研究表明，PDE11A mRNA在骨骼肌、前列腺、肾脏、肝脏、垂体、唾液腺和睾丸中含量最高。PDE11A至少表达为三个主要转录本，约为10.5。近似于8.5 kb，近似于6.0 kb，从而表明存在多个亚型。通过人体组织的PDE11A异构体的Western blotting检测到三种不同的蛋白，分别为大约78、大约65和大约56 kDa，进一步支持了这种可能性。重组人PDE11A1水解cGMP和cAMP， K-m值分别为0.52 μ M和1.04 μ M， V-max值相似。因此，PDE11A代表了一种双底物PDE，在生理条件下可能同时调节cGMP和cAMP。PDE11A对非选择性PDE抑制剂3-异丁基-1-甲基黄嘌呤（IBMX）以及通常被认为对cgmp选择性PDE相对特异性的抑制剂zaprinast和dipyridamole敏感，其IC50值分别为49.8 μ M、12.0 μ M和0.37 μ M。
We report here the cloning, expression, and characterization of human PDE11A1, a member of a distinct cyclic nucleotide phosphodiesterase (PDE) family. PDE11A exhibits less than or equal to 50% amino acid identity with the catalytic domains of all other PDEs, being most similar to PDE5, and has distinct biochemical properties. The human PDE11A1 cDNA isolated contains a complete open reading frame encoding a 490-amino acid enzyme with a predicted molecular mass of 55,786 Da. At the N terminus PDE11A1 has a single CAF domain homologous to that found in other signaling molecules, including PDE2, PDE5, PDE6, and PDE10, which constitutes a potential allosteric binding site for cGMP or another small ligand. Tissue distribution studies indicate that PDE11A mRNA occurs at highest levels in skeletal muscle, prostate, kidney, liver, pituitary, and salivary glands and testis. PDE11A is expressed as at least three major transcripts of approximate to 10.5. approximate to 8.5, and approximate to 6.0 kb, thus suggesting the existence of multiple subtypes. This possibility is further supported by the detection of three distinct proteins of approximate to 78, approximate to 65, and approximate to 56 kDa by Western blotting of human tissues for PDE11A isoforms. Recombinant human PDE11A1 hydrolyzes both cGMP and cAMP with K-m values of 0.52 mu M and 1.04 mu M, respectively, and similar V-max values. Therefore, PDE11A represents a dual-substrate PDE that may regulate both cGMP and cAMP under physiological conditions. PDE11A is sensitive to the nonselective PDE inhibitor 3-isobutyl-1-methylxanthine (IBMX) as well as zaprinast and dipyridamole, inhibitors that are generally considered relatively specific far the cGMP-selective PDEs, with IC50 values of 49.8 mu M, 12.0 mu M, and 0.37 mu M, respectively.