期刊
SCIENCE
卷 318, 期 5848, 页码 279-283出版社
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.1146113
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资金
- Cancer Research UK [11722] Funding Source: Medline
- NIGMS NIH HHS [GM065367, R01 GM065367] Funding Source: Medline
Despite the recent advances in single-molecule manipulation techniques, purely mechanical approaches cannot detect subtle conformational changes in the biologically important regime of weak forces. We developed a hybrid scheme combining force and fluorescence that allowed us to examine the effect of subpiconewton forces on the nanometer scale motion of the Holliday junction (HJ) at 100-hertz bandwidth. The HJ is an exquisitely sensitive force sensor whose force response is amplified with an increase in its arm lengths, demonstrating a lever-arm effect at the nanometer-length scale. Mechanical interrogation of the HJ in three different directions helped elucidate the structures of the transient species populated during its conformational changes. This method of mapping two-dimensional reaction landscapes at low forces is readily applicable to other nucleic acid systems and their interactions with proteins and enzymes.
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