Engineered Polypeptide Condensate Shifting for Molecular Concentration
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods lack effective compositions and methods for modulating molecules within cells to regulate cellular functions and maintain homeostasis, particularly in concentrating molecules for enzymatic reactions or signaling cascades.
Innovation Solution
An engineered polypeptide comprising a functional domain and a condensate shifting domain with an amino acid sequence of a nuclear receptor or fragment thereof, which forms a first condensate upon contact with a condensate modulator, allowing for liquid-liquid phase separation and controlled exchange of molecules, and can be shifted to a second condensate with different properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If molecules are concentrated in a specific location in a cell to initiate enzymatic reactions or signaling cascades, then cellular function regulation is improved, but the complexity of controlling molecular distribution and concentration increases
Solution Approach 1:
The patent uses condensate modulators as intermediary molecules that bind to the condensate shifting domain to control phase separation. These modulators act as mediators between the engineered polypeptide and the desired molecular concentration state, simplifying the control mechanism while achieving precise molecular distribution and concentration regulation.
Solution Approach 2:
The patent employs condensate modulators that induce phase separation by changing physical-chemical parameters such as concentration, temperature, or binding affinity. This allows dynamic control of molecular distribution and concentration within cells through reversible parameter changes, enabling enzymatic reactions and signaling cascades without complex control systems.
2Quantity of substance
If an engineered polypeptide forms a condensate to concentrate molecules, then molecular concentration and enzymatic activity are improved, but the ability to reverse or modulate the condensate formation becomes limited
Solution Approach 1:
The patent designs the engineered polypeptide with a condensate shifting domain that enables dynamic, reversible phase separation. The condensate modulators can induce or reverse condensate formation by binding to or dissociating from the polypeptide, allowing the system to adapt between different molecular concentration states. This dynamic control provides versatility in regulating molecular concentration for different cellular functions.
3Object-affected harmful factors
If modifications are made to the nuclear receptor sequence to decrease binding with nucleic acids or ligands, then off-target effects are reduced, but the binding affinity and effectiveness of the polypeptide may be compromised
Solution Approach 1:
The patent segments the nuclear receptor sequence into a condensate shifting domain that retains only the essential phase separation capability while removing or modifying regions responsible for nucleic acid and ligand binding. This segmentation allows the polypeptide to maintain its condensate-forming function and binding affinity for modulators while reducing off-target effects from unwanted interactions with cellular components.
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 engineered polypeptide effectively concentrates molecules, enhancing enzymatic activity by bringing molecules into close proximity and modulating their concentration, thereby regulating cellular processes and potentially treating diseases by administering the polypeptide or its encoding polynucleotide.
Implementation Method 1
the first condensate mediates liquid-liquid phase separation (LLPS)
Data Source
AI summary
Provided are compositions for modulating molecules in a cell. Also provided are methods for modulating molecules in a cell.