Qin Yue, Zhang Xiaoyu, Song Yanling, Zhong Bowen, Liu Lu, Wang Dongdong, Zhang Yahui, Lu Wenqi, Zhao Xinjia, Jia Zhiqi, Li Minmin, Zhang Lihua, Qing Guangyan
College of Pharmaceutical and Biological Engineering, Shenyang University of Chemical Technology No. 11 Street, Economic and Technological Development Zone Shenyang 110142 P. R. China.
CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences Dalian 116023 P. R. China
Chem Sci. 2023 Jul 13;14(31):8360-8368. doi: 10.1039/d3sc02140h. eCollection 2023 Aug 9.
SUMOylation is an important and highly dynamic post-translational modification (PTM) process of protein, and its disequilibrium may cause various diseases, such as cancers and neurodegenerative disorders. SUMO proteins must be accurately detected to understand disease states and develop effective drugs. Reliable antibodies against SUMO2/3 are commercially available; however, efficient detectors are yet to be developed for SUMO1, which has only 50% homology with SUMO2 and SUMO3. Here, using phage display technology, we identified two cyclic peptide (CP) sequences that could specifically bind to the terminal dodecapeptide sequence of SUMO1. Then we combined the CPs and polyethylene terephthalate conical nanochannel films to fabricate a nanochannel device highly sensitive towards the SUMO1 terminal peptide and protein; sensitivity was achieved by ensuring marked variations in both transmembrane ionic current and Faraday current. The satisfactory SUMO1-sensing ability of this device makes it a promising tool for the time-point monitoring of the SENP1 enzyme-catalyzed de-SUMOylation reaction and cellular imaging. This study not only solves the challenge of SUMO1 precise recognition that could promote SUMO1 proteomics analysis, but also demonstrates the good potential of the nanochannel device in monitoring of enzymes and discovery of effective drugs.
SUMO化是一种重要且高度动态的蛋白质翻译后修饰（PTM）过程，其失衡可能导致各种疾病，如癌症和神经退行性疾病。为了解疾病状态并开发有效药物，必须准确检测SUMO蛋白。市面上有可靠的抗SUMO2/3抗体；然而，针对与SUMO2和SUMO3仅有50%同源性的SUMO1，尚未开发出高效的检测方法。在此，我们利用噬菌体展示技术，鉴定出两个能特异性结合SUMO1末端十二肽序列的环肽（CP）序列。然后，我们将这些环肽与聚对苯二甲酸乙二醇酯锥形纳米通道膜相结合，制造出一种对SUMO1末端肽和蛋白质高度敏感的纳米通道装置；通过确保跨膜离子电流和法拉第电流都有显著变化来实现灵敏度。该装置令人满意的SUMO1传感能力使其成为用于监测SENP1酶催化的去SUMO化反应和细胞成像的时间点监测的有前途的工具。这项研究不仅解决了促进SUMO1蛋白质组学分析的SUMO1精确识别难题，还展示了纳米通道装置在酶监测和有效药物发现方面的良好潜力。
