Self-sacrificial tyrosine cleavage by an Fe:Mn oxygenase for the biosynthesis of para-aminobenzoate in Chlamydia trachomatis.
Self-sacrificial tyrosine cleavage by an Fe:Mn oxygenase for the biosynthesis of para-aminobenzoate in Chlamydia trachomatis.
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DOI：
10.1073/pnas.2210908119
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发表时间：
2022-09-27
影响因子：
11.1
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中科院分区：
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对氨基苯甲酸(PABA)作为必需的维生素四氢叶酸的前体，是合成核苷酸和氨基酸所必需的。沙眼衣原体是性传播感染的病原体，它缺乏规范的pabABC生物合成途径，而是利用一种新的策略来合成PABA，该策略是由衣原体蛋白相关死亡结构域(CADD)酶催化的。最初被指定为二铁酶，我们报告了高的体外活性依赖于铁和锰的化学计量添加。此外，CADD被证明激活O2以自我牺牲地从Tyr27合成PABA。这项工作解决了衣原体产生PABA的不寻常机制，这是开发治疗感染的治疗剂的主要目标。沙眼衣原体死亡结构域相关蛋白(CADD)参与了对氨基苯甲酸(PABA)的生物合成，对氨基苯甲酸是人类病原体沙眼衣原体生存和增殖所必需的叶酸辅因子。衣原体用于合成PABA的途径不同于大多数细菌使用的典型的多酶途径，该途径依赖于分支酸作为代谢前体。相反，最近的研究表明，CADD形成的PABA来自L酪氨酸。作为血红素加氧酶样二铁氧合酶(HDOS)超家族中的一员，CADD被提出使用二铁辅因子进行催化。然而，我们报道了CADD中最大的PABA形成发生在铁和锰的添加上，这意味着异质双金属Fe：Mn团簇是催化活性形式。同位素标记实验和蛋白质组学研究表明，CADD从蛋白质衍生的酪氨酸(Tyr27)产生PABA，酪氨酸是来自双金属位点的残基∼14？我们认为，这种自祭反应是通过一个可能的Fe：Mn原子团簇通过连接到“底物”Tyr的自由基继电器机制激活O2而发生的，然后是胺化和直接氧插入。这些结果为沙眼衣原体中PABA的形成提供了分子基础，为设计新的治疗药物提供了依据。
As a precursor to the essential vitamin tetrahydrofolate, para-aminobenzoate (pABA) is needed for nucleotide and amino acid synthesis. Chlamydia trachomatis, the causative agent of the sexually transmitted infection, lacks the canonical pabABC biosynthetic pathway and instead utilizes a novel stratagem for pABA synthesis that is catalyzed by the enzyme Chlamydia protein associating with death domains (CADD). Originally assigned as a diiron enzyme, we report high in vitro activity relies on the stoichiometric addition of both Fe and Mn. Furthermore, CADD is shown to activate O2 to self-sacrificially produce pABA from Tyr27. This work addresses the unusual mechanism by which Chlamydiae produce pABA, which is a prime target for the development of therapeutic agents to treat infections. Chlamydia protein associating with death domains (CADD) is involved in the biosynthesis of para-aminobenzoate (pABA), an essential component of the folate cofactor that is required for the survival and proliferation of the human pathogen Chlamydia trachomatis. The pathway used by Chlamydiae for pABA synthesis differs from the canonical multi-enzyme pathway used by most bacteria that relies on chorismate as a metabolic precursor. Rather, recent work showed pABA formation by CADD derives from l-tyrosine. As a member of the emerging superfamily of heme oxygenase–like diiron oxidases (HDOs), CADD was proposed to use a diiron cofactor for catalysis. However, we report maximal pABA formation by CADD occurs upon the addition of both iron and manganese, which implicates a heterobimetallic Fe:Mn cluster is the catalytically active form. Isotopic labeling experiments and proteomics studies show that CADD generates pABA from a protein-derived tyrosine (Tyr27), a residue that is ∼14 Å from the dimetal site. We propose that this self-sacrificial reaction occurs through O2 activation by a probable Fe:Mn cluster through a radical relay mechanism that connects to the “substrate” Tyr, followed by amination and direct oxygen insertion. These results provide the molecular basis for pABA formation in C. trachomatis, which will inform the design of novel therapeutics.
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影响因子：
64.8
作者：
Chatterjee, Abhishek;Abeydeera, N. Dinuka;Bale, Shridhar;Pai, Pei-Jing;Dorrestein, Pieter C.;Russell, David H.;Ealick, Steven E.;Begley, Tadhg P.
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Begley, Tadhg P.
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Nordlund, P
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2.9
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Jiang, Wei;Hoffart, Lee M.;Martin Bollinger, J., Jr.
通讯作者：
Martin Bollinger, J., Jr.
影响因子：
56.9
作者：
Jiang, Wei;Yun, Danny;Bollinger, J. Martin, Jr.
通讯作者：
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