一类 "新 "磷酸烯醇丙酮酸羧激酶的结构和功能差异--通过低聚物变化了解异构调节

M. Mcleod, Siddhi Balamurali, Robert Thorne, T. Holyoak
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摘要

磷酸烯醇丙酮酸羧激酶(PEPCK)是控制 TCA 循环的代谢酶。它们被认为是治疗糖尿病、癌症和结核分枝杆菌感染的潜在靶点,并在衰老和长寿中发挥作用。这些酶将草酰乙酸(OAA)转化为磷酸烯醇丙酮酸(PEP)。PEPCKs 广泛分布于生命体中,根据其催化反应中使用的磷酸供体的性质分为三类。在这三类PEPCKs中,研究最不深入的是依赖PPi的PEPCKs,它们在结构上和功能上都有别于使用核苷酸的PEPCKs(GTP和ATP)。依赖 PPi 的 PEPCKs 有一个保守的核心(约 60 kDa),它包含了依赖核苷酸类的一般折叠。然而,它们的结构有明显的附加部分(约 70 kDa),这些附加部分形成了异构位点和低聚物界面,很可能导致了不同的功能特性。在这里,我们利用尺寸排阻色谱法、酶动力学、晶体学和小角 X 射线散射来了解三种 PPi 依赖性 PEPCK 同工酶的结构和功能方面。发现伊斯拉放线菌(Actinomyces israelii)依赖 PPi 的 PEPCK 是一种组成型二聚体,其活性明显降低。弗氏丙酸杆菌(Pf-PPi-PEPCK)在酶催化反应、金属依赖性以及通过单体-二聚体转变的异源诱导活性调节方面与核苷酸同工酶不同。第三个同工酶来自人类寄生虫组织溶解恩塔米巴虫(Eh-PPi-PEPCK)。Eh-PPi-PEPCK 有三个具有不同序列的同工酶、
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Structural and functional divergence of a 'new' class of phosphoenolpyruvate carboxykinase – insights into allosteric regulation via oligomeric changes
Phosphoenolpyruvate carboxykinases (PEPCK) are metabolic enzymes controlling the TCA - cycle. They have been implicated as potential targets in treating diabetes, cancer, and Mycobacterium tuberculosis infections, and have a role in aging and longevity. These enzymes interconvert oxaloacetic acid (OAA) to form phosphoenolpyruvate (PEP). PEPCKs are widely distributed across life and occur in three classes depending on the nature of phosphoryl donor used for their catalyzed reactions. Of the three classes, the most understudied are PPi-dependent PEPCKs, which are structurally and functionally distinct from the nucleotide -using classes (GTP and ATP). PPi-dependent PEPCKs have a conserved core (~60 kDa) that comprises the general fold of both nucleotide-dependent classes. However, their structure has significant additions (~70 kDa) that form allosteric sites and oligomeric interfaces, and that have likely l ed to divergent functional properties. Here we have used size-exclusion chromatography, enzyme kinetics, crystallography and small-angle x-ray scattering to understand the structural and functional aspects of three PPi - dependent PEPCK isozymes. Actinomyces israelii PPi-dependent PEPCK is found as a constitutive dimer with significantly reduced activity. Propionibacterium freudenreichii (Pf - PPi-PEPCK) differs from its nucleotide counterparts in its enzyme - catalyzed reaction, metal - dependencies, and alloste rically induced activity - regulation via monomer - dimer transition. The third isozyme is from the human parasite Entamoeba histolytica (Eh-PPi-PEPCK). Eh-PPi-PEPCK occurs in three paralogs with different sequences,
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