Monomolecular Fluorescent Sensor for Protein Surface Tracking
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current fluorescent molecular sensors face challenges in tracking changes on specific protein surfaces due to high background signals in complex biochemical mixtures and within cells, and they often require significant conformational changes or labeling at specific positions, limiting their ability to detect protein modifications and binding interactions effectively.
Innovation Solution
Development of a monomolecular compound comprising a His-tag binder, a non-selective binder, and a fluorophore, where the His-tag binder selectively binds to His-tagged proteins, and the non-selective binder interacts with protein surfaces, enabling the detection of changes without requiring significant conformational changes or specific labeling, using a flexible linker and modifiable synthetic receptor to match different protein regions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescent molecular sensors utilize photo-induced electron transfer (PET), electronic energy transfer (EET), or fluorescence resonance energy transfer (FRET) processes to detect proteins, then detection sensitivity is improved, but background signal increases complicating use in complex biochemical mixtures and within cells
Solution Approach 1:
The patent extracts the fluorescent detection function from the protein binding function by using a separate fluorophore that binds to a peptide motif on the protein surface. This allows the sensor to detect protein presence and conformational changes without the harmful background signals associated with PET/EET/FRET processes in complex biochemical mixtures.
Solution Approach 2:
The patent introduces a peptide motif as an intermediary element that mediates between the fluorophore and the protein target. The fluorophore binds to this peptide motif, which is itself bound to the protein, creating a indirect detection mechanism that reduces background signal while maintaining detection sensitivity.
2Measurement precision
If genetically encoded fluorescent proteins (FPs) are used to track protein expression and localization, then spatial and temporal resolution is improved, but the large size of FPs interrupts the normal function of labeled proteins
Solution Approach 1:
The patent segments the detection function into two separate components: a small fluorophore that binds to a peptide motif on the protein surface, rather than using a large fluorescent protein fusion. This segmentation allows the protein to maintain its native function while still enabling detection with high spatial and temporal resolution.
Solution Approach 2:
The patent uses a small, simple fluorophore that can be quickly synthesized and applied, replacing the need for complex, large fluorescent protein constructs. This small molecule approach maintains detection capability while minimizing interference with protein function.
3Adaptability or versatility
If small molecule-based fluorescent probes bind to short peptide motifs on proteins, then protein labeling in complex biological environments is enabled, but the ability to detect protein modifications and binding interactions is limited without significant conformational changes or specific labeling
Solution Approach 1:
The patent makes the sensor system dynamic by allowing the fluorophore-peptide motif complex to respond to protein conformational changes and binding interactions. The sensor can detect these dynamic changes in protein structure and interactions, providing precise measurement of protein modifications without requiring significant conformational changes or specific labeling.
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
This approach allows for the sensitive detection of protein surface modifications and binding interactions with reduced background noise, enabling the tracking of changes on specific proteins within complex environments without interfering with the protein's normal function.
Implementation Method 1
a fluorophore, which emits fluorescent light when excited
Implementation Method 2
The His-tag binder is represented by the structure of formula D... M is a metal ion
Data Source
Figure 1~1c
Figure 2~2c
Figure 3
AI summary
This invention is directed to monomolecular sensors, comprising a selective binder, a non-selective binder and a fluorophore, which can track changes that occur on the surface of labeled proteins. This invention is further directed to the use of such sensors for identifying binding partners of specific proteins. This invention is further directed to His-tag binding compounds and uses thereof in the preparation of genetically targeted detectable molecules and sensors which can specifically bind tag-labeled proteins.