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.

AFS Protein Folding

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.

AFS Protein Folding Talin Distance-Time Curve

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

AFS Example Data Protein Folding Equilibrium Dyamics

3 Protein unfolding equilibrium dynamics at a constant force.

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

Solutions

AFS™ Parallel Single-Molecule Force Spectroscopy
AFS™

Parallel Single-Molecule Force Spectroscopy

Acoustic Force Spectrosocpy (AFS™) is a new single-molecule and single-cell manipulation method capable of applying acoustic forces on hundreds of biomolecules in parallel for precise experimentation with high throughput. It enables scientists to probe thousands of individual molecules in parallel (such as RNA, DNA, proteins, and living cells), allowing statistical analysis of the mechanical properties of biological properties of biological systems based on a single experiment.

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