Leveraging Substrate Promiscuity of a Radical S-Adenosyl-L-methionine RiPP Maturase toward Intramolecular Peptide Cross-Linking Applications.
Leveraging Substrate Promiscuity of a Radical S-Adenosyl-L-methionine RiPP Maturase toward Intramolecular Peptide Cross-Linking Applications.
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利用自由基 S-腺苷-L-甲硫氨酸 RiPP 成熟酶的底物混杂性进行分子内肽交联应用。
DOI：
10.1021/acscentsci.2c00501
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发表时间：
2022-08-24
影响因子：
18.2
通讯作者：
Bandarian, Vahe
中科院分区：
文献类型：
--
作者：
Eastman, Karsten A. S.;Kincannon, William M.;Bandarian, Vahe
自由基S-腺苷-L-甲硫氨酸（RS）酶作用于多种底物，并催化广泛的复杂自由基介导的转化。自由基非α-碳硫醚肽（ranthipeptides）是一类由核糖体合成并经后修饰的肽（RiPPs）。RS酶PapB催化位于六个Cys-X3-Asp/Glu基序中的Cys/Asp（或Cys/Glu）残基之间的硫醚交联的形成。在这份报告中，使用一个最小的基板，包含一个单一的交联基序，我们探索的PapB的基板范围，并表明该酶是高度混杂的，并会接受各种Cys-Xn-Asp序列，其中n = 0-6。此外，我们表明，该酶将独立地在含有两个来自野生型序列的基序的肽序列中引入内联和嵌套的硫醚交联。此外，该酶接受在Cys或Asp位置含有d-氨基酸的肽。利用这些观察结果来产生FDA批准的治疗剂奥曲肽的硫醚环化类似物，其中Cys-Glu交联取代药物中发现的二硫化物。这些发现突出了PapB显著的底物耐受性，并显示了RS RiPP成熟酶在生物技术应用中的实用性。PapB修饰多种肽底物。PapB的混杂性被用于体外合成FDA批准的药物奥曲肽的硫醚大环化类似物。
Radical S-adenosyl-l-methionine (RS) enzymes operate on a variety of substrates and catalyze a wide range of complex radical-mediated transformations. Radical non-α-carbon thioether peptides (ranthipeptides) are a class of ribosomally synthesized and post-translationally modified peptides (RiPPs). The RS enzyme PapB catalyzes the formation of thioether cross-links between Cys/Asp (or Cys/Glu) residues located in six Cys-X3-Asp/Glu motifs. In this report, using a minimal substrate that contains a single cross-link motif, we explore the substrate scope of the PapB and show that the enzyme is highly promiscuous and will accept a variety of Cys-Xn-Asp sequences where n = 0–6. Moreover, we show that the enzyme will introduce in-line and nested thioether cross-links independently in peptide sequences that contain two motifs derived from the wild-type sequence. Additionally, the enzyme accepts peptides that contain d-amino acids at either the Cys or the Asp position. These observations are leveraged to produce a thioether cyclized analogue of the FDA-approved therapeutic agent octreotide, with a Cys-Glu cross-link replacing the disulfide that is found in the drug. These findings highlight the remarkable substrate tolerance of PapB and show the utility of RS RiPP maturases in biotechnological applications. PapB modifies a variety of peptide substrates. The promiscuity of PapB was leveraged toward the in vitro synthesis of a thioether macrocyclized analogue of the FDA-approved drug octreotide.
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影响因子：
5.5
作者：
Benjdia A;Balty C;Berteau O
通讯作者：
Berteau O
影响因子：
18.2
作者：
Burkhart BJ;Kakkar N;Hudson GA;van der Donk WA;Mitchell DA
通讯作者：
Mitchell DA
影响因子：
4.8
作者：
Crain, Adam V.;Broderick, Joan B.
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Broderick, Joan B.
影响因子：
16.6
作者：
King AM;Anderson DA;Glassey E;Segall-Shapiro TH;Zhang Z;Niquille DL;Embree AC;Pratt K;Williams TL;Gordon DB;Voigt CA
通讯作者：
Voigt CA
影响因子：
2.9
作者：
deGruyter JN;Malins LR;Baran PS
通讯作者：
Baran PS