Genetic Fusion of Thermoresponsive Polypeptides with UCST‐type Behavior Mediates 1D Assembly of Coiled‐Coil Bundlemers
Genetic Fusion of Thermoresponsive Polypeptides with UCST‐type Behavior Mediates 1D Assembly of Coiled‐Coil Bundlemers
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温敏多肽与 UCST 型行为的基因融合介导卷曲线圈捆绑器的一维组装
DOI：
10.1002/anie.202301331
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Kiick, Kristi L.
中科院分区：
文献类型：
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作者：
Patkar, Sai S.;Tang, Yao;Bisram, Arriana M.;Zhang, Tianren;Saven, Jeffery G.;Pochan, Darrin J.;Kiick, Kristi L.
不同长度的热响应性弹性蛋白样多肽（RLP）被遗传融合到两种不同的计算设计的具有不同热稳定性的卷曲卷曲肽中，通过温度触发的RLP单元相分离来开发新的策略来组装卷曲卷曲肽。通过凝胶电泳、质谱和氨基酸分析验证了它们在细菌表达宿主中的成功生产。圆二色（CD）光谱、紫外-可见（UV/Vis）比浊法和动态光散射（DLS）测量证实了卷曲线圈的稳定性，并表明rlp的热敏相行为在遗传融合的杂交多肽中得到了保留。低温透射电子显微镜和粗粒度模型显示，用热响应性rlp功能化卷曲线圈导致其热触发的非共价组装成纳米纤维组装。
Thermoresponsive resilin‐like polypeptides (RLPs) of various lengths were genetically fused to two different computationally designed coiled coil‐forming peptides with distinct thermal stability, to develop new strategies to assemble coiled coil peptides via temperature‐triggered phase separation of the RLP units. Their successful production in bacterial expression hosts was verified via gel electrophoresis, mass spectrometry, and amino acid analysis. Circular dichroism (CD) spectroscopy, ultraviolet‐visible (UV/Vis) turbidimetry, and dynamic light scattering (DLS) measurements confirmed the stability of the coiled coils and showed that the thermosensitive phase behavior of the RLPs was preserved in the genetically fused hybrid polypeptides. Cryogenic‐transmission electron microscopy and coarse‐grained modeling revealed that functionalizing the coiled coils with thermoresponsive RLPs leads to their thermally triggered noncovalent assembly into nanofibrillar assemblies.