Song Wen-Wen, Qian Zhi-Gang, Liu Hao, Chen Hai-Feng, Kaplan David L, Xia Xiao-Xia
State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, 800 Dongchuan Road, Shanghai 200240, China.
Department of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, Massachusetts 02155, United States.
ACS Macro Lett. 2021 Apr 20;10(4):395-400. doi: 10.1021/acsmacrolett.1c00062. Epub 2021 Mar 9.
Despite considerable progress having been made in thermosensitive protein hydrogels, regulating their thermal transitions remains a challenge due to the intricate molecular structures and interactions of the underlying protein polymers. Here we report a genetic fusion strategy to tune the unique dual thermal transitions of the C-terminal domain (CTD) of spider major ampullate spidroin 1, and explore the regulation mechanism by biophysical characterization and molecular dynamics simulations. We found that the fusion of elastin-like polypeptides (ELPs) tuned the dual transition temperatures of CTD to a physiologically relevant window, by introducing extra hydrogen bonding at low temperatures and hydrophobic interactions at high temperatures. The resulting hydrogels constructed from the fusion proteins were demonstrated to be a promising vehicle for cell preservation and delivery. This study provides insights on the regulation of the dual thermosensitive protein hydrogels and suggests a potential application of the hydrogels for consolidated cell storage and delivery.
尽管在热敏蛋白水凝胶方面已经取得了相当大的进展，但由于潜在蛋白质聚合物复杂的分子结构和相互作用，调节它们的热转变仍然是一个挑战。在此，我们报告了一种基因融合策略，用于调节蜘蛛主要壶腹蛛丝蛋白1 C末端结构域（CTD）独特的双热转变，并通过生物物理表征和分子动力学模拟探索其调节机制。我们发现，弹性蛋白样多肽（ELP）的融合通过在低温下引入额外的氢键和在高温下引入疏水相互作用，将CTD的双转变温度调节到生理相关窗口。由融合蛋白构建的所得水凝胶被证明是一种用于细胞保存和递送的有前景的载体。这项研究为双热敏蛋白水凝胶的调节提供了见解，并暗示了水凝胶在巩固细胞储存和递送方面的潜在应用。
