FEATHER

FEATHER automates identification of unfolding and unbinding events in single-molecule force spectroscopy (SMFS) force-extension data to deduce rupture forces, loading rates, and related biophysical parameters.

Key Features:

• Automation: Automatically locates unfolding and unbinding events within SMFS records without requiring prior knowledge of the system under study.
• Precision and Accuracy: Improves event location precision by 30-fold, enhances accuracy of loading rate and rupture force distributions by eightfold, and reduces false positives threefold versus reference algorithms.
• Scalability: Implements a linear algorithm that scales efficiently with large SMFS datasets.
• User-defined Parameters: Exposes two user-defined parameters that control event detection while avoiding bias toward dominant behaviors.

Scientific Applications:

• Protein dynamics and energetics: Enables accurate deduction of rupture force and loading rate to derive zero-force dissociation rate constants (k_o) and distances to transition states (Δx‡).
• Protein–ligand interactions: Characterizes binding energetics via rupture force and loading rate analysis from SMFS experiments.
• Nucleic acid structures: Identifies unfolding/unbinding events in nucleic acid SMFS to probe structural stability and mechanics.
• Rupture force distribution analysis: Analyzes the shape of rupture force distributions to infer energy barrier heights (ΔG‡).

Methodology:

Analyzes SMFS force-extension curves recorded during constant-velocity probe retraction, automatically identifies unfolding/unbinding events, deduces rupture forces and loading rates, and operates with linear algorithmic scaling.