Structural and Biochemical Analysis of the Essential Diadenylate Cyclase CdaA from Listeria monocytogenes*
Structural and Biochemical Analysis of the Essential Diadenylate Cyclase CdaA from Listeria monocytogenes*
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DOI：
10.1074/jbc.m114.630418
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发表时间：
2015-01
期刊：
影响因子：
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通讯作者：
J. Rosenberg;A. Dickmanns;P. Neumann;K. Gunka;J. Arens;V. Kaever;J. Stülke;R. Ficner;F. Commichau
中科院分区：
文献类型：
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作者：
J. Rosenberg;A. Dickmanns;P. Neumann;K. Gunka;J. Arens;V. Kaever;J. Stülke;R. Ficner;F. Commichau
背景：单核增生李斯特菌CdaA是一种必需的二腺苷酸环化酶。结果：CdaA活性依赖于锰、钴离子。结论：CdaA对金属辅助因子的要求不同寻常。意义：必需酶的鉴定对开发新型抗生素具有重要意义。最近发现的第二信使环二磷酸腺苷（c-二磷酸腺苷）参与了几个重要的细胞过程，如细胞壁代谢、DNA完整性的维持、离子运输、转录调节和酶功能的变构调节。有趣的是，c-di-AMP对革兰氏阳性模型细菌枯草芽孢杆菌的生长至关重要。虽然枯草芽孢杆菌的基因组编码三种可以在功能上相互替代的c-二腺苷酸环化酶，但与系统发育相关的人类病原体，如单核增生李斯特菌和金黄色葡萄球菌，只有一种酶，即二腺苷酸环化酶CdaA。由于CdaA对这些细菌的生长也是必不可少的，因此该酶是开发新型抗生素的有希望的靶标。在这里，我们提出了在许多人类病原体中保守的单核增生乳杆菌CdaA二腺苷酸环化酶结构域的第一个晶体结构。此外，环化酶的生化表征显示了不同寻常的金属辅因子需求。
Background: Listeria monocytogenes CdaA is an essential diadenylate cyclase. Results: CdaA activity depends on manganese and cobalt ions. Conclusion: CdaA has an unusual requirement for metal cofactors. Significance: Characterization of essential enzymes is important for developing novel antibiotics. The recently identified second messenger cyclic di-AMP (c-di-AMP) is involved in several important cellular processes, such as cell wall metabolism, maintenance of DNA integrity, ion transport, transcription regulation, and allosteric regulation of enzyme function. Interestingly, c-di-AMP is essential for growth of the Gram-positive model bacterium Bacillus subtilis. Although the genome of B. subtilis encodes three c-di-AMP-producing diadenlyate cyclases that can functionally replace each other, the phylogenetically related human pathogens like Listeria monocytogenes and Staphylococcus aureus possess only one enzyme, the diadenlyate cyclase CdaA. Because CdaA is also essential for growth of these bacteria, the enzyme is a promising target for the development of novel antibiotics. Here we present the first crystal structure of the L. monocytogenes CdaA diadenylate cyclase domain that is conserved in many human pathogens. Moreover, biochemical characterization of the cyclase revealed an unusual metal cofactor requirement.