Selection for constrained peptides that bind to a single target protein.
Selection for constrained peptides that bind to a single target protein.
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DOI：
10.1038/s41467-021-26350-4
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发表时间：
2021-11-03
影响因子：
16.6
通讯作者：
Voigt CA
中科院分区：
文献类型：
--
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
肽类次级代谢产物在自然界中很常见，具有多种药理学相关功能，从抗生素到跨界信号传导。在这里，我们提出了一种方法来设计大肠杆菌中的大型修饰肽库，并在体内筛选它们，以确定那些结合到一个单一的目标感兴趣的。使用从微生物RiPP（核糖体合成和后修饰肽）途径收集的修饰的酶产生受约束的肽支架，并使其多样化以构建大型文库。RiPP与蛋白质靶标的结合导致内含肽催化的RNA聚合酶σ因子的释放，其驱动选择性标记的表达。作为概念验证，进行了结合SARS-CoV-2刺突受体结合结构域的选择。发现1625 Da限制性肽（AMK-1057）与ACE 2衍生肽的结合亲和力相似（990 ± 5 nM）。这证明了一种可推广的方法来鉴定粘附于单个蛋白质靶标的约束肽，作为朝向用于治疗和诊断的“分子胶”的一步。肽类次级代谢产物具有多种功能。在这里，作者提出了一种方法来设计和筛选一个大型的修饰肽库，在E。大肠杆菌针对感兴趣的靶标。
Peptide secondary metabolites are common in nature and have diverse pharmacologically-relevant functions, from antibiotics to cross-kingdom signaling. Here, we present a method to design large libraries of modified peptides in Escherichia coli and screen them in vivo to identify those that bind to a single target-of-interest. Constrained peptide scaffolds were produced using modified enzymes gleaned from microbial RiPP (ribosomally synthesized and post-translationally modified peptide) pathways and diversified to build large libraries. The binding of a RiPP to a protein target leads to the intein-catalyzed release of an RNA polymerase σ factor, which drives the expression of selectable markers. As a proof-of-concept, a selection was performed for binding to the SARS-CoV-2 Spike receptor binding domain. A 1625 Da constrained peptide (AMK-1057) was found that binds with similar affinity (990 ± 5 nM) as an ACE2-derived peptide. This demonstrates a generalizable method to identify constrained peptides that adhere to a single protein target, as a step towards “molecular glues” for therapeutics and diagnostics. Peptide secondary metabolites have a diverse range of functions. Here the authors present a method to design and screen a large library of modified peptides in E. coli against a target of interest.
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影响因子：
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作者：
Gavrish E;Sit CS;Cao S;Kandror O;Spoering A;Peoples A;Ling L;Fetterman A;Hughes D;Bissell A;Torrey H;Akopian T;Mueller A;Epstein S;Goldberg A;Clardy J;Lewis K
通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Benard, Francois