Prevention of Protease-Induced Degradation of Desmoplakin via Small Molecule Binding

TL;DR

Researchers found small molecules that can prevent the degradation of desmoplakin, a protein linked to heart disease, by blocking protease activity.

Contribution

The study identifies drug-like molecules that block protease-induced degradation of desmoplakin without affecting protease function.

Findings

Several small molecules prevent degradation of wildtype and disease-variant desmoplakin in the presence of trypsin and calpain.

Molecular dynamics suggest that long hydrophobic molecules bind in a shallow groove near the cleavage site.

These findings provide a foundation for developing pharmacological treatments for hypersensitive desmoplakin variants.

Abstract

Desmoplakin (DSP) is a large (~260 kDa) protein found in the desmosome, the subcellular structure that links the intermediate filament network of one cell to its neighbor. A mutation “hot-spot” within the NH2-terminal of the DSP protein (residues 299–515) is associated with arrhythmogenic cardiomyopathy. In a subset of DSP variants, disease is linked to calpain hypersensitivity. Previous studies show that calpain hypersensitivity can be corrected in vitro through the addition of a bulky residue neighboring the cleavage site, suggesting that physically blocking calpain accessibility is a viable strategy to restore DSP levels. Here, we aim to find drug-like molecules that also block calpain-dependent degradation of DSP. To do this, we screened ~2500 small molecules to identify compounds that specifically rescue DSP protein levels in the presence of proteases. We find that several molecules, including sodium dodecyl sulfate, palmitoylethanolamide, GW0742, salirasib, eprosarten mesylate, and GSK1838705A prevent wildtype and disease-variant-carrying DSP protein degradation in the presence of both trypsin and calpain without altering protease function. Computational screenings did not predict which molecules would protect DSP, likely due to a lack of specific DSP–drug interactions. Molecular dynamic simulations of DSP–drug complexes suggest that some long hydrophobic molecules can bind in a shallow hydrophobic groove that runs alongside the protease cleavage site. Identification of these compounds lays the groundwork for pharmacological treatment for individuals harboring these hypersensitive DSP variants.

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