Subnanometer-Precision Measurements of Transmembrane Motions of Biomolecules in Plasma Membranes Using Quenchers in Extracellular Environment.

Wenqing Hou, Dongfei Ma, Xiaolong He, Weijing Han, Jianbing Ma, Hao Wang, Chunhua Xu, Ruipei Xie, Qihui Fan, Fangfu Ye, Shuxin Hu, Ming Li, Ying Lu

Nano letters 2021 Jan 13

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Characterization of biomolecular dynamics at cellular membranes lags far behind that in solutions because of challenges to measure transmembrane trafficking with subnanometer precision. Herein, by introducing nonfluorescent quenchers into extracellular environment of live cells, we adopted Förster resonance energy transfer from one donor to multiple quenchers to measure positional changes of biomolecules in plasma membranes. We demonstrated the method by monitoring flip-flops of individual lipids and by capturing transient states of the host defense peptide LL-37 in plasma membranes. The method was also applied to investigate the interaction of the necroptosis-associated protein MLKL with plasma membranes, showing a few distinct depths of MLKL insertion. Our method is especially powerful to quantitate the dynamics of proteins at the cytosolic leaflets of plasma membranes which are usually not accessible by conventional techniques. The method will find wide applications in the systematic analysis of fundamental cellular processes at plasma membranes.

Citation

Wenqing Hou, Dongfei Ma, Xiaolong He, Weijing Han, Jianbing Ma, Hao Wang, Chunhua Xu, Ruipei Xie, Qihui Fan, Fangfu Ye, Shuxin Hu, Ming Li, Ying Lu. Subnanometer-Precision Measurements of Transmembrane Motions of Biomolecules in Plasma Membranes Using Quenchers in Extracellular Environment. Nano letters. 2021 Jan 13;21(1):485-491