The function of proteins is determined by a combination of their structure and dynamics. Acquisition of structural information has recently been revolutionized by advances in electron microscopy, however protein dynamics is difficult to obtain over long timescales. Here we show a novel plasmon ruler based single molecule approach to study the conformational dynamics of single proteins over 24 hours at video rate in and out of equilibrium (i. e. in the absence and presence of ATP). This approach does not impose external force on the proteins and allows to measure protein dynamics over several orders of magnitude, which cannot be achieved by the established single molecule techniques. We used this technique to explore the dynamics of the  heat shock protein 90 (Hsp90) and find (beside the well-known fast dynamics in the range netween 0.1 and 10 s) states with very long dwell times (linked to rarely visited states) in the minute timescale, which could indicate, for instance, pathways to protein denaturation. Our plasmon ruler based method will enable to access the full complexity of correlated local and global conformational dynamics on the level of individual proteins and therefore extend and complement the current "structure / function" paradigm with a novel "structure / timescale / function" description. Access to the dynamics of a single protein over more than 6 orders of magnitude further allows us to address important questions like conformational heterogeneity among proteins, ergodic behavior, and nonMarkovian dynamics, ultimately extending the often static view of biomolecules with a more dynamic picture .
We use plasmon rulers to follow the conformational dynamics of a single protein for up to 24 h at a video rate. The plasmon ruler consists of two gold nanospheres connected by a single protein linker. In our experiment, we follow the dynamics of the molecular chaperone heat shock protein 90 , which is known to show open and closed conformations. Our measurements confirm the previously known conformational dynamics with transition times in the second to minute time scale and reveals new dynamics on the time scale of minutes to hours. Plasmon rulers thus extend the observation bandwidth 3 / 4 orders of magnitude with respect to single-molecule fluorescence resonance energy transfer and enable the study of molecular dynamics with unprecedented precision .
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The function of proteins is determined by a combination of their structure and dynamics. Acquisition of structural information has recently been revolutionized by advances in electron microscopy, however protein dynamics is difficult to obtain over long timescales. Here we show a novel plasmon ruler based single molecule approach to study
We use plasmon rulers to follow
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single proteins over
a single protein for up to
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hours at video rate in and out of equilibrium (i. e. in the absence and presence of ATP). This approach does not impose external force on the proteins and allows to measure protein dynamics over several orders of magnitude, which cannot be achieved by the established single molecule techniques. We used this technique to explore
h at a video rate. The plasmon ruler consists of two gold nanospheres connected by a single protein linker. In our experiment, we follow
A
molecular chaperone
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(Hsp90) and find (beside the well-known fast dynamics in the range netween 0.1 and 10 s) states with very long dwell times (linked to rarely visited states) in the minute timescale, which could indicate, for instance, pathways to protein denaturation. Our plasmon ruler based method will enable to access the full complexity of correlated local and global conformational
, which is known to show open and closed conformations. Our measurements confirm the previously known conformational dynamics with transition times in the second to minute time scale and reveals new
R
level of individual proteins and therefore extend and complement the current "structure
time scale of minutes to hours. Plasmon rulers thus extend the observation bandwidth 3
R
function" paradigm with a novel "structure / timescale / function" description. Access to the dynamics of a single protein over more than 6
4
R
further allows us to address important questions like conformational heterogeneity among proteins, ergodic behavior, and nonMarkovian dynamics, ultimately extending the often static view of biomolecules with a more dynamic picture
with respect to single-molecule fluorescence resonance energy transfer and enable the study of molecular dynamics with unprecedented precision