Characterization of Plasmodium falciparum cGMP-dependent protein kinase (PfPKG):: Antiparasitic activity of a PKG inhibitor
Characterization of Plasmodium falciparum cGMP-dependent protein kinase (PfPKG):: Antiparasitic activity of a PKG inhibitor
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DOI：
10.1016/j.molbiopara.2005.10.020
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发表时间：
2006-03-01
影响因子：
1.5
通讯作者：
Liberator, PA
中科院分区：
文献类型：
--
作者：
Diaz, CA;Allocco, J;Liberator, PA
环GMP依赖性蛋白激酶（PKG）已在弓形虫中被生物化学和遗传学验证为负责三取代吡咯4-[2-（2-氨基苯基）-3-甲基-2-氧代-4-氧代-2-氧代-3-氧代-2-（4-氟苯基）-5-（1-甲基哌啶-4-基）-1H-吡咯-3-基]吡啶（化合物1）[Biftu T，Feng D，Ponpipom M，等人，作为新型抗球虫剂的寄生虫cGMP依赖性蛋白激酶的2，3-二芳基吡咯抑制剂的合成和SAR。Bioorg Med Chem Lett 2005;15：3296-301; Gurnett AM，Liberator PA，Dulski PM，et al. Purification and molecular characterization of cGMP-dependent protein kinase from Apicomplexan parasites.一种新的化疗靶点。J Biol Chem 2002; 277：15913-22; Donald RGK，Allocco J，Singh SB，et al.弓形虫环GMP依赖性激酶：一种必需寄生虫蛋白激酶的化学靶向。真核细胞2002; 1：317-28; Nare B，Allocco J，Liberator PA，Donald RGK.评价环GMP依赖性蛋白激酶抑制剂治疗鼠弓形虫病：γ-干扰素是有效的。AntimicrobAgents Chemother 2002;46：300-7]。化合物I抑制顶复门（Apicomplexa）亚门的几种相关原生动物寄生虫的生长。已通过cGMP亲和和MonoQ离子交换层析从恶性疟原虫（PfPKG）中部分纯化了天然PKG活性。使用抗PKG抗血清检测的富集98 kDa蛋白的生化级分含有cGMP诱导的蛋白激酶活性，其对化合物1的抑制敏感。为了使PJPKG更彻底的表征，我们表达了一个合成的cDNA纳入弓形虫密码子偏好（Pf（Tg）PKG）在弓形虫寄生虫。纯化的重组Pf（Tg）PKG的蛋白激酶活性受到环鸟苷酸的刺激，希尔系数为2，具有显着的协同性。化合物1对Pf（Tg）PKG激酶活性的底物偏好和抑制均与天然PJPKG以及来自艾美耳球虫属的PKG酶相似。和弓形虫。我们的结论是PfPKG具有类似于以前的特点apicomplexan PKG酶的生化和药理学特性。化合物1对体外培养的恶性疟原虫的血细胞阶段具有活性。在伯氏疟原虫感染的小鼠模型中，化合物1延迟寄生虫血症的发作，但不能治愈寄生虫感染。(c)2005 Elsevier B. V.保留所有权利。
Cyclic GMP-dependent protein kinase (PKG) has been biochemically and genetically validated in Toxoplasma gondii as a primary target responsible for the antiparasitic activity of the trisubstituted pyrrole 4-[2-(4-fluorophenyl)-5-(1-methylpiperidine-4-yl)-1H pyrrol-3-yl] pyridine (Compound 1) [Biftu T, Feng D, Ponpipom M, et al. Synthesis and SAR of 2,3-diarylpyrrole inhibitors of parasite cGMP-dependent protein kinase as novel anticoccidial agents. Bioorg Med Chem Lett 2005;15:3296-301; Gurnett AM, Liberator PA, Dulski PM, et al. Purification and molecular characterization of cGMP-dependent protein kinase from Apicomplexan parasites. A novel chemotherapeutic target. J Biol Chem 2002;277:15913-22; Donald RGK, Allocco J, Singh SB, et al. Toxoplasma gondii cyclic GMP-dependent kinase: Chemotherapeutic targeting of an essential parasite protein kinase. Eukaryotic Cell 2002; 1: 317-28; Nare B, Allocco J, Liberator PA, Donald RGK. Evaluation of a cyclic GMP-dependent protein kinase inhibitor in treatment of murine Toxoplasmosis: Gamma interferon is required for efficacy. Antimicrob Agents Chemother 2002;46:300-7]. Compound I inhibits the growth of several related protozoan parasites of the subphylum Apicomplexa. Native PKG activity has been partially purified by cGMP-affinity and MonoQ ion exchange chromatography from Plasmodium falciparum (PfPKG). Biochemical fractions enriched for a 98 kDa protein detected using anti-PKG antisera, contain cGMP-induced protein kinase activity that is sensitive to inhibition by Compound 1. To enable a more thorough characterization of PJPKG we expressed a synthetic cDNA incorporating T gondii codon preference (Pf(Tg)PKG) in T gondii parasites. The protein kinase activity of purified recombinant Pf(Tg)PKG is stimulated by cGMP, with significant cooperativity as demonstrated by a Hill coefficient of 2. Both substrate preference and inhibition of Pf(Tg)PKG kinase activity by Compound 1 are similar to that seen with native PJPKG, as well as PKG enzymes from Eimeria spp. and T gondii. We conclude that PfPKG has biochemical and pharmacological properties that are similar to previously characterized apicomplexan PKG enzymes. Compound 1 is active against blood cell stages of P. falciparum cultured in vitro. In a Plasmodium berghei mouse model of infection, Compound 1 delays the onset of parasitemia but does not cure the parasite infection. (c) 2005 Elsevier B.V. All rights reserved.