Measure Protein Folding and Conformational Dynamics with High Throughput

Part of: Protein Folding and Conformational Changes

Multiplexed study of protein folding & conformational dynamics

In this experiment, multiple proteins are tethered between a bead and a glass surface using DNA handles on each side. Pulling the beads towards the acoustic node causes the different protein domains to unfold.

Figure 1 shows multiple force-distance curves obtained by a single Talin protein submitted to alternating stretching and relaxation cycles using AFS. Figure 2 displays an enlarged snippet of an individual force-distance curve corresponding to a single pulling cycle. Here, while ramping the force from 15 to 19 pN, we observe a series of four unfolding events — corresponding to four individual protein domains —ranging between 30 and 100 nm. The unfolding events can be clearly distinguished owing to the high-resolution distance measurement capability of this technology.

Figure 3 shows an illustration of a typical protein unfolding experiment performed at a constant force. Equilibrium dynamics show the transition between different intermediate states. AFS has the ability to measure equilibrium dynamics because the intrinsic force clamp drives the piezo at a constant voltage. Additionally, AFS allows measuring many molecules in parallel, which boosts the experimental data throughput.

1 Force-distance curves representing multiple stretching cycles of the same individual Talin protein.

2 Zoom- in of an individual force-distance curve corresponding to a single pulling cycle, covering a force ramp range of 4 pN.

3 Protein unfolding equilibrium dynamics at a constant force.

Sample and data courtesy of Prof. Yan Jie at the National University of Singapore.

Solutions

C-Trap® Optical Tweezers Fluorescence & Label-free Microscopy
C-Trap

Optical Tweezers and Fluorescence Microscopy

The C-Trap is the world’s first instrument that allows simultaneous manipulation and visualization of molecular interactions in real-time. It combines high-resolution optical tweezers, confocal microscopy or STED nanoscopy with an advanced microfluidics system in a truly integrated and correlated solution.

M-Trap® Optical Tweezers Fluorescence & Label-free Microscopy
m-Trap

Optical Tweezers

The m-Trap is the first entry-level optical tweezers instrument specifically developed for high-resolution single-molecule research. Ultra-high force resolution and stability, with incredible throughput, ease of use and modularity ‒ all at an unprecedented price level.

Key Product FeaturesC-Trap®m-Trap®
Conformational changes and states
Force-induced structural transitions
Visualization of ligand interactions using fluorescence
Investigation of higher order structures using FRET
Rapid buffer exchange for fast experimental workflow

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