Recombinant production of rhesus θ-defensin-1 (RTD-1) using a bacterial expression system.
Recombinant production of rhesus θ-defensin-1 (RTD-1) using a bacterial expression system.
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DOI：
10.1039/c2mb05451e
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发表时间：
2012-04
影响因子：
--
通讯作者：
Camarero JA
中科院分区：
文献类型：
--
作者：
Gould A;Li Y;Majumder S;Garcia AE;Carlsson P;Shekhtman A;Camarero JA
防御素是一种在哺乳动物先天免疫防御中起重要作用的抗菌肽。与哺乳动物α-和β-防御素不同，恒河猴θ防御素-1（RTD-1）仅包含由三个二硫键和不寻常的骨架环状拓扑结构稳定的18个氨基酸。本文首次报道了利用细菌表达系统重组表达完全折叠的θ-防御素RTD-1。这是使用分子内天然化学连接结合修饰的蛋白质剪接单元来完成的。RTD-1在体外或体内产生。估计RTD-1的细胞内产生达到104 μM的细胞内浓度。重组RTD-1被证明是正确折叠的，其特征在于异核NMR和其特异性抑制致死因子蛋白酶的能力。折叠的θ-防御素的重组生产打开了基于这种肽支架产生肽库的可能性，所述肽库可用于开发细胞内筛选和定向进化技术。
Defensins are antimicrobial peptides that are important in the innate immune defense of mammals. In contrast to mammalian α- and β-defensins, rhesus theta defensin-1 (RTD-1) comprises only 18 amino acids stabilized by three disulfide bonds and an unusual backbone cyclic topology. In this work we report for the first time the recombinant expression of the fully folded θ-defensin RTD-1 using a bacterial expression system. This was accomplished using an intramolecular native chemical ligation in combination with a modified protein-splicing unit. RTD-1 was produced either in vitro or in vivo. In-cell production of RTD-1 was estimated to reach an intracellular concentration of ≈ 4 μM. Recombinant RTD-1 was shown to be correctly folded as characterized by heteronucelar-NMR and by its ability to specifically inhibit Lethal Factor protease. The recombinant production of folded θ-defensins opens the possibility to produce peptide libraries based on this peptide scaffold that could be used to develop in-cell screening and directed evolution technologies.
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DOI：
10.1111/j.1399-3011.2004.00155.x
发表时间：
2004-06-01
期刊：
JOURNAL OF PEPTIDE RESEARCH
影响因子：
--
作者：
Owen, SM;Rudolph, D;Lal, RB
通讯作者：
Lal, RB
DOI：
10.1073/pnas.96.24.13703
发表时间：
1999-11-23
影响因子：
11.1
作者：
Bessette, PH;Åslund, F;Georgiou, G
通讯作者：
Georgiou, G
影响因子：
56.9
作者：
Tang, YQ;Yuan, J;Selsted, ME
通讯作者：
Selsted, ME
影响因子：
16.6
作者：
Kimura, RH;Tran, AT;Camarero, JA
通讯作者：
Camarero, JA
影响因子：
2.9
作者：
Kimura, Richard H.;Steenblock, Erin R.;Camarero, Julio A.
通讯作者：
Camarero, Julio A.