文献类型：期刊文献

英文题名：T232K/K238Q Aerolysin Nanopore for Mapping Adjacent Phosphorylation Sites of a Single Tau Peptide

作者：Li, Shuang[1,2];Wu, Xue-Yuan[2];Li, Meng-Yin[1];Liu, Shao-Chuang[1];Ying, Yi-Lun[1];Long, Yi-Tao[1]

机构：[1]Nanjing Univ, Sch Chem & Chem Engn, State Key Lab Analyt Chem Life Sci, Nanjing 210023, Peoples R China;[2]East China Univ Sci & Technol, Sch Chem & Mol Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2020

卷号：4

期号：11

外文期刊名：SMALL METHODS

收录：;EI(收录号：20202108692495);WOS:【SCI-EXPANDED(收录号:WOS:000530371900001)】；

基金：S.L. and X.-Y.W. contributed equally to this work. This research was supported by the National Natural Science Foundation of China (21922405, 61871183, and 21834001). Y.-L.Y. is sponsored by National Ten Thousand Talent Program for young top-notch talent. The authors thank Prof. Jin Huang and Ya-Qian Wang from East China University of Science and Technology for their help with the aerolysin site-directed mutagenesis; Prof. Qun Xu from Renji Hospital, Shanghai Jiao Tong University for the helpful discussions on phosphorylated tau fragments.

语种：英文

外文关键词：aerolysin nanopores; phosphorylation sites; single-molecule analyses; tau peptides

摘要：Tau phosphorylation shows direct clinical importance as the hyperphosphorylation and aggregation of tau exists in a range of tauopathies. However, it is still challenging to study tau phosphorylation owing to its multiple and adjacent phosphorylation sites in the tau sequence. To address this challenge, here, a designed T232K/K238Q mutant aerolysin nanopore is introduced which synergistically incorporates the enhanced electrostatic interaction at T232K site and the high repelling barrier at K238Q site. The distinct current blockages produced by a T232K/K238Q aerolysin sensor achieve nearly 100% identification accuracy for the characteristic distribution of unphosphorylated tau peptide, pS262-, pT263-tau peptide, and pS262/pT263-tau peptide. The excellent sensing ability of the T232K/K238Q nanopore originates from the extremely slow translocation speed which prolongs the duration up to tens or hundreds milliseconds for a nine-amino-acid peptide. It is envisioned that the design ideas of the T232K/K238Q aerolysin nanopore can be further applied to analyze other protein/peptide post-translational modification as it provides the exquisite sensitivity for identifying the modification of single amino acids.