Lin Senzhu, Chen Gefei, Meng Qing
Institute of Biological Sciences and Biotechnology, Donghua University, Shanghai 201620, China.
Center for Alzheimer Research, Karolinska Institute, Stockholm 14157, Sweden.
Sheng Wu Gong Cheng Xue Bao. 2016 Dec 25;32(12):1704-1714. doi: 10.13345/j.cjb.160157.
To provide technical support for spider silk functional modification, we developed a simple and efficient functional platform via intein trans-splicing. Small ubiquitin-related modifier protein (SUMO) was fused to the recombinant spider silk protein (W2CT) by peptide bond via S0 split intein Ssp DnaB trans-splicing, resulting in a protein SUMOW2CT. However, incorporation of exogenous protein led to mechanical property defect and lower fiber yield, and also slowed down the fiber assembly velocity but no obvious differences in supercontraction and chemical resistance when compared with fibers from W2CT (W). SUMO protease digestion showed positive results on the fibers, indicating that the SUMO protein kept its native conformation and bioactive. Above all, this work provides a technical support for spider silk high simply and efficient functionalized modification.
为了为蜘蛛丝功能修饰提供技术支持，我们通过内含肽反式剪接开发了一个简单高效的功能平台。小泛素相关修饰蛋白（SUMO）通过S0分裂内含肽Ssp DnaB反式剪接通过肽键与重组蜘蛛丝蛋白（W2CT）融合，产生了蛋白SUMOW2CT。然而，外源蛋白的掺入导致了机械性能缺陷和纤维产量降低，并且还减慢了纤维组装速度，但与来自W2CT（W）的纤维相比，在超收缩和耐化学性方面没有明显差异。SUMO蛋白酶消化在纤维上显示出阳性结果，表明SUMO蛋白保持了其天然构象和生物活性。最重要的是，这项工作为蜘蛛丝的简单高效功能化修饰提供了技术支持。
