Otopetrin Proton Channels: Patch-Clamp and Fluorescence Detection
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Solution Overview
Problem
Proton channels that can transport protons into eukaryotic cells have not been molecularly identified, limiting the understanding and modulation of proton translocation activity.
Innovation Solution
Identify and characterize otopetrin polypeptides, such as Otop1, Otop2, and Otop3, as proton-selective ion channels, and develop methods to assess their modulation through proton translocation activity using techniques like patch clamp and fluorescence assays.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If otopetrin polypeptides are identified and characterized as proton-selective ion channels, then understanding and modulation of proton translocation activity is improved, but the complexity of identifying and characterizing these channels increases
Solution Approach 1:
The patent uses intermediary substances such as fluorescent dyes (e.g., pHrodo red, BCECF) and electrophysiological recording techniques as mediators to detect and characterize proton translocation activity. These intermediaries translate the biological function of otopetrin channels into measurable signals, enabling reliable identification without requiring direct observation of proton movement.
Solution Approach 2:
The patent replaces direct mechanical or chemical measurement of proton translocation with optical detection methods (fluorescence assays) and electrical measurement methods (patch clamp). This substitution allows for non-invasive, real-time monitoring of channel activity while reducing the complexity of direct proton measurement.
2Adaptability or versatility
If methods to assess modulation of proton translocation activity are developed, then targeted modulation for therapeutic applications is enabled, but the complexity of assessment increases
Solution Approach 1:
The patent develops assessment methods using universal techniques such as patch clamp electrophysiology and fluorescence imaging that can be applied to multiple otopetrin channel variants (Otop1, Otop2, Otop3) and multiple modulator types. These multi-functional methods enable simultaneous assessment of different channels and compounds using the same experimental platform, enhancing therapeutic adaptability.
Solution Approach 2:
The patent utilizes parameter changes in fluorescent dyes (pH-sensitive fluorescence intensity) and electrical properties (membrane potential, current amplitude) to assess modulator effects. By monitoring these measurable parameters before and after modulator application, the patent enables quantitative assessment of therapeutic modulation while maintaining relatively simple assessment protocols.
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
Provides a means to identify modulators of otopetrin-mediated proton translocation, enabling targeted modulation of proton channels for therapeutic applications.
Implementation Method 1
the transmembrane protein Otopetrin 1 (Otop1) and certain related transmembrane proteins are identified as proton-selective ion channels
Implementation Method 2
determining a proton translocation activity mediated by the otopetrin polypeptide
Implementation Method 3
determining a proton translocation activity mediated by the otopetrin polypeptide, or the functional portion thereof
Data Source
Figure 1a~1h
Figure 2a~2g
Figure 3a~3f
AI summary
Presented herein are compositions and devices comprising otopetrin polypeptides and uses thereof for identifying modulators of proton translocation activity through an otopetrin polypeptide, or functional portion thereof.