RiPP enzyme heterocomplex structure-guided discovery of a bacterial borosin α-N-methylated peptide natural product.
RiPP enzyme heterocomplex structure-guided discovery of a bacterial borosin α-N-methylated peptide natural product.
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DOI：
10.1039/d3cb00093a
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发表时间：
2023-10-04
影响因子：
4.1
通讯作者：
中科院分区：
文献类型：
--
作者：
酰胺肽主链甲基化是一种特征性翻译后修饰，存在于称为 borosins 的核糖体合成和翻译后修饰肽天然产物 (RiPP) 家族中。此前，我们通过生物信息学方法鉴定了细菌中超过 1500 个假定的硼素途径；然而，没有任何途径与已知的次生代谢物相关。通过对 Shewanella oneidensis MR-1 中的 borosin 途径进行深入表征，我们现已鉴定出一种细菌来源的 borosin 天然产物，名为 Shewanellamide A。通过创建和分析一系列前体变体以及对变体前体和甲基转移酶复合物的晶体学询问，促进了 Borosin 的鉴定。在分析来自 S. oneidensis 的两种蛋白酶的同时，通过比较 S. oneidensis 敲除和过表达菌株，预测并确认了代谢物蛋白水解成熟的可能边界。总而言之，S. oneidensis 天然产物被发现是一种 16 聚体线性肽，具有两个主链甲基化，使 Shewanellamide A 成为迄今已鉴定的少数硼松素代谢物之一，也是第一个来自细菌的代谢物。通过结合体外结构-活性测定、晶体学和体内敲除实验，我们鉴定并表征了第一种细菌硼松天然产物。
Amide peptide backbone methylation is a characteristic post-translational modification found in a family of ribosomally synthesized and post-translationally modified peptide natural products (RiPPs) called borosins. Previously, we bioinformatically identified >1500 putative borosin pathways in bacteria; however, none of the pathways were associated with a known secondary metabolite. Through in-depth characterization of a borosin pathway in Shewanella oneidensis MR-1, we have now identified a bacterially derived borosin natural product named Shewanellamide A. Borosin identification was facilitated by the creation and analysis of a series of precursor variants and crystallographic interrogation of variant precursor and methyltransferase complexes. Along with assaying two proteases from S. oneidensis, probable boundaries for proteolytic maturation of the metabolite were projected and confirmed via comparison of S. oneidensis knockout and overexpression strains. All in all, the S. oneidensis natural product was found to be a 16-mer linear peptide featuring two backbone methylations, establishing Shewanellamide A as one of the few borosin metabolites yet identified, and the first from bacteria. Through a combination of in vitro structure–activity assays, crystallography, and in vivo knockout experiments, we have identified and structurally characterized the first bacterial borosin natural product.
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影响因子：
15
作者：
Beck, Johannes G.;Chatterjee, Jayanta;Kessler, Horst
通讯作者：
Kessler, Horst
使用N-甲基化杂色氨基酸的工程β-折叠。
影响因子：
8.4
作者：
Ghosh D;Lahiri P;Verma H;Mukherjee S;Chatterjee J
通讯作者：
Chatterjee J
影响因子：
4.4
作者：
Gao H;Wang X;Yang ZK;Palzkill T;Zhou J
通讯作者：
Zhou J
影响因子：
3.7
作者：
Han S;Machhi S;Berge M;Xi G;Linke T;Schoner R
通讯作者：
Schoner R
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K