HD-[HD-GYP] Phosphodiesterases: Activities and Evolutionary Diversification within the HD-GYP Family.
HD-[HD-GYP] Phosphodiesterases: Activities and Evolutionary Diversification within the HD-GYP Family.
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DOI：
10.1021/acs.biochem.0c00257
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发表时间：
2020-06-30
期刊：
影响因子：
2.9
通讯作者：
Pandelia ME
中科院分区：
文献类型：
--
作者：
Sun S;Pandelia ME
环状二核苷酸是调节许多过程的信号分子，包括免疫应答和毒力因子产生。它们在细菌中的细胞水平通过金属依赖性磷酸二酯酶（即EAL和HD-GYP蛋白）进行微调，其中HD-GYP属于较大的HD结构域超家族。在这项研究中，我们首先专注于HD-[HD-GYP]亚家族的催化性能和金属离子和底物的范围，由两个HD域组成。我们将SO 3491鉴定为VCA 0681的同源物和HD-[HD-GYP]的第二个实例。这两种蛋白质水解c-di-GMP和3′3′ c-GAMP，并与各种金属离子配位，但只有Fe和较小程度的Co支持水解。这些蛋白质仅在二价铁形式下具有活性，而在单电子更氧化的FeIIFeIII状态下不具有活性。虽然C-末端HD-GYP结构域对于活性是必需的，但N-末端HD结构域的作用仍然未知。我们发现，N-末端的网站是重要的蛋白质的稳定性，影响个人的表观kcat和KM（但不是kcat/KM），并不能结合c-di-GMP，从而排除其参与环二核苷酸传感。我们继续进行系统发育分析，以检查HD-[HD-GYP]与其余HD-GYP的分布和功能关系。该基因组学提供了一个相关图，绘制了HD-GYPs的进化和功能多样化之间的联系，作为预测金属辅因子的化学性质，活性水平和反应结果的模板。
Cyclic dinucleotides are signaling molecules that modulate many processes, including immune response and virulence factor production. Their cellular levels in bacteria are fine-tuned by metal-dependent phosphodiesterases, namely, the EAL and HD-GYP proteins, with HD-GYPs belonging to the larger HD domain superfamily. In this study, we first focus on the catalytic properties and the range of metal ions and substrates of the HD-[HD-GYP] subfamily, consisting of two HD domains. We identified SO3491 as a homologue of VCA0681 and the second example of an HD-[HD-GYP]. Both proteins hydrolyze c-di-GMP and 3′3′ c-GAMP and coordinate various metal ions, but only Fe and to a lesser extent Co support hydrolysis. The proteins are active only in the diferrous form and not in the one-electron more oxidized FeIIFeIII state. Although the C-terminal HD-GYP domain is essential for activity, the role of the N-terminal HD domain remains unknown. We show that the N-terminal site is important for protein stability, influences the individual apparent kcat and KM (but not kcat/KM), and cannot bind c-di-GMP, thus precluding its involvement in cyclic dinucleotide sensing. We proceeded to perform phylogenetic analyses to examine the distribution and functional relationships of the HD-[HD-GYP]s to the rest of the HD-GYPs. The phylogeny provides a correlation map that draws a link between the evolutionary and functional diversification of HD-GYPs, serving as a template for predicting the chemical nature of the metallocofactor, level of activity, and reaction outcome.
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影响因子：
6.7
作者：
Jones CJ;Utada A;Davis KR;Thongsomboon W;Zamorano Sanchez D;Banakar V;Cegelski L;Wong GC;Yildiz FH
通讯作者：
Yildiz FH
影响因子：
3.6
作者：
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DOI：
10.1016/j.bbapap.2015.04.015
发表时间：
2015-08
影响因子：
3.2
作者：
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通讯作者：
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影响因子：
64.8
作者：
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通讯作者：
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影响因子：
2.9
作者：
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通讯作者：
Kurtz DM Jr