Biosynthesis of Isonitrile Lipopeptide Metallophores from Pathogenic Mycobacteria.
Biosynthesis of Isonitrile Lipopeptide Metallophores from Pathogenic Mycobacteria.
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病原分枝杆菌异腈脂肽金属载体的生物合成。
DOI：
10.1021/acs.biochem.2c00611
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发表时间：
2023
期刊：
影响因子：
2.9
通讯作者：
Zhang,Wenjun
中科院分区：
文献类型：
--
作者：
DelRioFlores,Antonio;Narayanamoorthy,Maanasa;Cai,Wenlong;Zhai,Rui;Yang,Siyue;Shen,Yuanbo;Seshadri,Kaushik;DeMatias,Kyle;Xue,Zhaoqiang;Zhang,Wenjun
异腈脂肽（INLP）是已知的致病性分枝杆菌的毒力介导的金属转运，但其生物合成仍然是模糊的。在这项工作中，我们使用体外生物化学测定，定点诱变，化学合成和光谱技术，以审查所需的INLP生物合成在分枝杆菌的核心酶的活性。与环境链霉菌相比，致病性分枝杆菌INLP的生物合成具有相似的化学逻辑和酶机制，主要区别在于脂肪酰基链的长度，这一过程受多个酶的控制。我们对非血红素铁（II）和α-酮戊二酸依赖性双加氧酶（包括结核分枝杆菌（Mtb）的Rv 0097）的深入研究表明，它识别一个独立的小分子底物，不同于最近的假设，即Mtb中Rv 0097需要载体蛋白。Rv 0097中的一个关键残基被进一步鉴定为决定链霉菌和分枝杆菌之间不同的脂肪酰链长度特异性。
Isonitrile lipopeptides (INLPs) are known to be related to the virulence of pathogenic mycobacteria by mediating metal transport, but their biosynthesis remains obscure. In this work, we use in vitro biochemical assays, site-directed mutagenesis, chemical synthesis, and spectroscopy techniques to scrutinize the activity of core enzymes required for INLP biosynthesis in mycobacteria. Compared to environmentalStreptomyces, pathogenicMycobacteriumemploy a similar chemical logic and enzymatic machinery in INLP biosynthesis, differing mainly in the fatty-acyl chain length, which is controlled by multiple enzymes in the pathway. Our in-depth study on the non-heme iron(II) and α-ketoglutarate-dependent dioxygenase for isonitrile generation, including Rv0097 fromMycobacterium tuberculosis(Mtb), demonstrates that it recognizes a free-standing small molecule substrate, different from the recent hypothesis that a carrier protein is required for Rv0097 inMtb. A key residue in Rv0097 is further identified to dictate the varied fatty-acyl chain length specificity betweenStreptomycesandMycobacterium.