RyR2 Channel Stabilization via Cryo-EM Ligand Binding
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Inherited mutations and stress-induced post-translational modifications in the ryanodine receptor (RyR) lead to calcium leaks, causing various diseases such as skeletal myopathies, heart failure, and exercise-induced sudden death, for which existing treatments are inadequate.
Innovation Solution
A composition comprising a complex suspended in a solid medium, specifically vitreous ice, containing a protein and a synthetic compound, where the protein is a ryanodine receptor 2 protein (RyR2) or its mutant, along with methods for predicting ligand binding and identifying compounds with RyR2 modulatory activity using cryo-EM and computational analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If RyR2 channel is left unmodified, then normal calcium release function is maintained, but calcium leaks occur due to mutations and post-translational modifications causing disease
Solution Approach 1:
The patent uses small molecule compounds as intermediaries to bind to the RyR2 channel and modulate its function. These compounds act as mediators between the mutated/post-modified RyR2 channel and the desired functional state, stabilizing the closed conformation and preventing calcium leaks without requiring genetic modification or complex therapeutic interventions
Solution Approach 2:
The patent employs compounds that induce parameter changes in the RyR2 channel by altering its conformational equilibrium. The binding of these compounds changes the energy landscape of the channel, shifting the balance toward the closed state and modifying the channel's gating parameters to prevent pathological calcium release
2Measurement precision
If cryo-EM structure determination is performed on RyR2 complexes, then high-resolution binding site information is obtained, but sample preparation and analysis complexity increases
Solution Approach 1:
The patent employs preliminary action in sample preparation by pre-stabilizing the RyR2 channel in a specific conformational state through compound binding before flash-freezing. This preliminary stabilization ensures that the channel maintains its functional state during the cryo-EM process, enabling high-resolution structure determination without requiring complex post-processing or reconstruction techniques
Solution Approach 2:
The patent creates composite structures by forming stable complexes between the RyR2 channel and small molecule compounds. These composite materials exhibit enhanced structural stability and defined conformations that are ideal for cryo-EM analysis, allowing researchers to obtain high-resolution binding site information while simplifying the overall structural determination process
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The solution effectively stabilizes the closed state of the RyR2 channel, preventing calcium leaks and restoring normal function, thereby addressing disease symptoms and improving treatment outcomes for associated conditions.
Implementation Method 1
a composition comprising a complex suspended in a solid medium, the solid medium comprising vitreous ice
Data Source
AI summary
The present disclosure relates to methods and compositions useful for the identification of a ryanodine receptor modulator binding site in ryanodine receptor type 2 (RyR2). The present disclosure also provides compositions useful for the analysis of the ryanodine receptor modulator binding site in RyR2 via cryo-EM. The present disclosure further provides computational methods for identifying compounds that bind to RyR2.