Calmodulin-dependent cyclic nucleotide phosphodiesterase (PDE1) is a pharmacological target of differentiation-inducing factor-1, an antitumor agent isolated from Dictyostelium
Calmodulin-dependent cyclic nucleotide phosphodiesterase (PDE1) is a pharmacological target of differentiation-inducing factor-1, an antitumor agent isolated from Dictyostelium
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DOI：
10.1158/0008-5472.can-03-3551
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发表时间：
2004-04-01
期刊：
影响因子：
11.2
通讯作者：
Kubohara, Y
中科院分区：
文献类型：
--
作者：
Shimizu, K;Murata, T;Kubohara, Y
分化诱导因子-1（DIF-1）是从盘基网柄藻（Dictyosteelanddiscoideum）中分离的一种有效的抗增殖剂，其在体外诱导哺乳动物细胞的生长停滞和分化。然而，DIF-1的特异性靶分子尚未鉴定。在本研究中，我们试图在哺乳动物细胞中鉴定DIF-1的靶分子，检测DIF-1及其类似物对一些候选酶活性的影响。在K562白血病细胞中，10-40 μ m剂量依赖性地抑制细胞生长并增加细胞内环AMP浓度。然后发现0.5-20 μ m的DIF-1在体外以剂量依赖性方式抑制钙调蛋白（CaM）依赖性环核苷酸磷酸二酯酶（PDE 1）。动力学分析表明，DIF-1作为一个竞争性抑制剂的PDE 1对底物环AMP。由于DIF-1没有显著影响其他PDE或CaM依赖性酶的活性，此外，DIF-1的异构体是一种效力较低的抑制剂，因此我们得出结论，PDE 1是DIF-1的药理学和特异性靶点。
The differentiation-inducing factor-1 (DIF-1)isolated from Dictyostelium discoideum is a potent antiproliferative agent that induces growth arrest and differentiation in mammalian cells in vitro. However, the specific target molecule(s) of DIF-1 has not been identified. In this study, we have tried to identify the target molecule(s) of DIF-1 in mammalian cells, examining the effects of DIF-1 and its analogs on the activity of some candidate enzymes. DIF-1 at 10-40 mum dose-dependently suppressed cell growth and increased the intracellular cyclic AMP concentration in K562 leukemia cells. It was then found that DIF-1 at 0.5-20 mum inhibited the calmodulin (CaM)-dependent cyclic nucleotide phosphodiesterase (PDE1) in vitro in a dose-dependent manner. Kinetic analysis revealed that DIF-1 acted as a competitive inhibitor of PDE1 versus the substrate cyclic AMP. Because DIF-1 did not significantly affect the activity of other PDEs or CaM-dependent enzymes and, in addition, an isomer of DIF-1 was a less potent inhibitor, we have concluded that PDE1 is a pharmacological and specific target of DIF-1.