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Mechanosensitive channel MscL gating transitions coupling with constriction point shift

Protein Sci. 2024-04; 
Mingfeng Zhang, Siyang Tang, Xiaomin Wang, Sanhua Fang, Yuezhou Li
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Proteins, Expression, Isolation and Analysis … Histag anti-body (A00186, Genscript) was used to determine the MscL protein. … MscL protein was then added to lipid vesicles at a protein-to-lipid mass ratio of 1:100 for reconstitution. … Get A Quote

摘要

The mechanosensitive channel of large conductance (MscL) acts as an "emergency release valve" that protects bacterial cells from acute hypoosmotic stress, and it serves as a paradigm for studying the mechanism underlying the transduction of mechanical forces. MscL gating is proposed to initiate with an expansion without opening, followed by subsequent pore opening via a number of intermediate substates, and ends in a full opening. However, the details of gating process are still largely unknown. Using in vivo viability assay, single channel patch clamp recording, cysteine cross-linking, and tryptophan fluorescence quenching approach, we identified and characterized MscL mutants with different occupancies of con... More

关键词

MscL, constriction point, gating pore, gating substate, gating transitions, mechanosensitive channel, protien-lipid interaction