Phage display and selection of lanthipeptides on the carboxy-terminus of the gene-3 minor coat protein.
Phage display and selection of lanthipeptides on the carboxy-terminus of the gene-3 minor coat protein.
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DOI：
10.1038/s41467-017-01413-7
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发表时间：
2017-11-15
影响因子：
16.6
通讯作者：
Prassler J
中科院分区：
文献类型：
--
作者：
Urban JH;Moosmeier MA;Aumüller T;Thein M;Bosma T;Rink R;Groth K;Zulley M;Siegers K;Tissot K;Moll GN;Prassler J
核糖体合成和后修饰肽（Ribosomally synthesized and post-deletionally modified peptides，RIPPs）是一类具有药物样性质的天然产物。为了充分利用RIPP作为肽药物候选物的潜力，需要用于其系统工程的工具。在这里，我们报告的工程lanthipeptides，一个子类的RIPPs的特点是多个硫醚循环，酶促引入的区域和立体特异性的方式，通过噬菌体展示。这是通过异源共表达线性羊毛硫肽前体与来自远亲细菌的修饰酶一起沿着融合到被广泛忽视的噬菌体M13次要外壳蛋白pIII的C-末端，而不是常规使用的N-末端来实现的。我们观察到，C-末端前体肽融合到pIII的酶促修饰的生产细胞的细胞质中，随后显示为成熟的环肽的噬菌体表面上。大的C-末端展示文库的生物淘选容易地鉴定对尿激酶纤溶酶原激活物（uPA）和链霉亲和素特异的人工羊毛硫肽配体。羊毛硫肽是一类具有潜在药物样性质的环状后修饰肽。在这里，作者通过在大肠杆菌中表达羊毛硫肽前体作为与噬菌体M13外壳蛋白pIII的C末端融合物来开发噬菌体展示系统。大肠杆菌沿着的异源修饰酶。
Ribosomally synthesized and post-translationally modified peptides (RiPPs) are an emerging class of natural products with drug-like properties. To fully exploit the potential of RiPPs as peptide drug candidates, tools for their systematic engineering are required. Here we report the engineering of lanthipeptides, a subclass of RiPPs characterized by multiple thioether cycles that are enzymatically introduced in a regio- and stereospecific manner, by phage display. This was achieved by heterologous co-expression of linear lanthipeptide precursors fused to the widely neglected C-terminus of the bacteriophage M13 minor coat protein pIII, rather than the conventionally used N-terminus, along with the modifying enzymes from distantly related bacteria. We observe that C-terminal precursor peptide fusions to pIII are enzymatically modified in the cytoplasm of the producing cell and subsequently displayed as mature cyclic peptides on the phage surface. Biopanning of large C-terminal display libraries readily identifies artificial lanthipeptide ligands specific to urokinase plasminogen activator (uPA) and streptavidin. Lanthipeptides are a class of cyclic post-translationally modified peptides with potential drug-like properties. Here the authors develop a phage display system by expressing lanthipeptide precursors as C-terminal fusions to the phage M13 coat protein pIII in E. coli along with the heterologous modifying enzymes.
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影响因子：
15
作者：
Liu, Wei;Chan, Alice S. H.;Vederas, John C.
通讯作者：
Vederas, John C.
DOI：
10.1073/pnas.0913677107
发表时间：
2010-06-08
影响因子：
11.1
作者：
Li, Bo;Sher, Daniel;van der Donk, Wilfred A.
通讯作者：
van der Donk, Wilfred A.
影响因子：
4.8
作者：
Lubelski, Jacek;Khusainov, Rustem;Kuipers, Oscar P.
通讯作者：
Kuipers, Oscar P.
影响因子：
8
作者：
Kapust, RB;Waugh, DS
通讯作者：
Waugh, DS
影响因子：
3.7
作者：
Férir G;Petrova MI;Andrei G;Huskens D;Hoorelbeke B;Snoeck R;Vanderleyden J;Balzarini J;Bartoschek S;Brönstrup M;Süssmuth RD;Schols D
通讯作者：
Schols D