中国科技论文在线-应用与环境生物学报.DOCVIP

PAGE FILENAME gaojianmoban PAGE Foldon介导的寡聚化类弹性蛋白多肽的相变特性葛慧华 张丹丹 吴姝羽 邱 岳 张光亚**华侨大学化工学院生物工程与技术系 厦门 361021摘 要 为了解SpyCatcher-ELPs40相变异常的原因并考察寡聚化结构域对其相变行为的影响，本文将Foldon（F）分别与SpyCatcher-ELPs40（C-E）和ELPs40（E）融合，构建了拓扑结构为非共价三臂星型寡聚化的C-E-F和E-F；探究了不同盐浓度和类型对其相变特性的影响，并与其它ELPs进行比较。结果发现：融合了Foldon的ELPs40的相变温度高于线型ELPs120而低于线型ELPs40及SpyTag/SpyCatcher介导的共价三臂星型ELPs，这说明ELPs拓扑结构会影响其相变行为。非共价三臂星型C-E-F的相变温度变化幅度较大，与等重复数的C-E和ELPs40相比，平均分别高出28.75 ℃和35.6 ℃，尤其在NaCl浓度为0.8 mol/L时，相差分别为41.2 ℃和47.1 ℃。同时，Na2CO3浓度在0.7 mol/L及以上时，非共价三臂星型C-E-F才会在低温下发生相变，而其在较低浓度的Na2SO4溶液中相变现象明显，不符合Hofmeister效应，是首次报道的非线型ELPs特殊的相变行为。这主要与Foldon和SpyCatcher的表面电荷以及寡聚态形式有关。（图6 表1 参32）关键词 类弹性蛋白多肽；相变温度；拓扑结构与性能；Hofmeister离子序；蛋白寡聚化CLC Q615、Q51、Q71收稿日期 Received: 2017-**-** 接受日期 Accepted: 20**-**-***国家自然科学基金项目资助 Supported by the National Natural Science Foundation of China .**通讯作者 Corresponding author (E-mail: zhgyghh@ )The phase transition behavior of elastin-like polypeptides oligomerized with the Foldon domainGE Huihua，ZHANG Dandan，WU Shuyu, QIU Yue，ZHANG Guangya**Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 361021, ChinaAbstractObjectives: To investigate the underlying mechanism of the unique phase transition behavior of SpyCatcher-ELPs40 (C-E) and the influences of oligomerization domain on the phase transition of C-E.Methods: We constructed a non-covalent three-armed star oligomerization of C-E-F and E-F by fusing Foldon (F) domain with SpyCatcher-ELPs40 (C-E) and ELPs40 (E). Results: Results showed that the phase transition temperature of ELPs40 fused with Foldon was higher than that of ELPs120, while lower than those of ELPs40 and the SpyTag/SpyCatcher-mediated covalent three-armed ELPs. This indicated that the topology of ELPs could affect its phase transition behavior. The phase transition temperature of C-E-F was 28.8℃ and 35.6 ℃ higher than those of C-E and ELPs40, although C-E-F had similar sequence with C-E and ELPs40. Especially, when the concentration of