B12-dependent photoresponsive protein hydrogels for controlled stem cell/protein release
B12-dependent photoresponsive protein hydrogels for controlled stem cell/protein release
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DOI：
10.1073/pnas.1621350114
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发表时间：
2017-06-06
影响因子：
11.1
通讯作者：
Sun, Fei
中科院分区：
文献类型：
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作者：
Wang, Ri;Yang, Zhongguang;Sun, Fei
由于对其结构和功能特性的精确控制，基因工程蛋白质水凝胶已成为生物医学应用的有希望的候选者。鉴于对创建刺激响应“智能”水凝胶的需求不断增长，我们在这里展示了通过使用基因编码的 SpyTag-SpyCatcher 化学在温和的生理条件下共价聚合腺苷钴胺素 (AdoB(12)) 依赖性光感受器 C 端腺苷钴胺素结合域 (CarH(C)) 蛋白质来合成完全基于蛋白质的光响应水凝胶。由此产生的由物理自组装CarH(C)聚合物组成的水凝胶在光照下表现出快速的凝胶-溶胶转变，这使得能够从3D培养物中轻松释放/回收3T3成纤维细胞和人间充质干细胞(hMSC)，同时保持其活力。共价交联的 CarH(C) 水凝胶还被设计为以光依赖性方式封装和释放大体积的球状蛋白，例如 mCherry。将刺激响应蛋白直接组装成水凝胶代表了设计动态可调材料的通用策略。
Thanks to the precise control over their structural and functional properties, genetically engineered protein-based hydrogels have emerged as a promising candidate for biomedical applications. Given the growing demand for creating stimuli-responsive "smart" hydrogels, here we show the synthesis of entirely protein-based photoresponsive hydrogels by covalently polymerizing the adenosylcobalamin (AdoB(12))-dependent photoreceptor C-terminal adenosylcobalamin binding domain (CarH(C)) proteins using genetically encoded SpyTag-SpyCatcher chemistry under mild physiological conditions. The resulting hydrogel composed of physically self-assembled CarH(C) polymers exhibited a rapid gel-sol transition on light exposure, which enabled the facile release/recovery of 3T3 fibroblasts and human mesenchymal stem cells (hMSCs) from 3D cultures while maintaining their viability. A covalently cross-linked CarH(C) hydrogel was also designed to encapsulate and release bulky globular proteins, such as mCherry, in a light-dependent manner. The direct assembly of stimuli-responsive proteins into hydrogels represents a versatile strategy for designing dynamically tunable materials.