Phytochrome Domain Light Control Protein Interaction
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Solution Overview
Problem
Current systems for controlling protein interactions within cells rely heavily on chemical effectors that require diffusion, which can be toxic and lack spatial and temporal precision, while light-regulated systems face limitations in reversibility and specificity.
Innovation Solution
The use of phytochrome domains and interacting peptides that convert between red and far-red light states, allowing for spatial and temporal regulation of protein interactions by light exposure, with specific binding to the Pfr state, enabling reversible and detectable protein-protein interactions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If chemical effectors are used to control protein interactions, then the control can be achieved within the cell, but the system suffers from toxicity and lacks spatial and temporal precision
Solution Approach 1:
The patent replaces chemical effector systems with a light-based optical system to control protein interactions. The phytochrome domain acts as a photosensor that converts light signals into conformational changes, enabling non-toxic, spatially and temporally precise control of protein-protein interactions without the harmful effects of chemical diffusion into the cell
Solution Approach 2:
The patent introduces a phytochrome domain as an intermediary component that mediates between light signals and protein interaction control. This intermediary converts external light stimuli into intracellular conformational changes, enabling precise control while avoiding direct introduction of toxic chemical effectors into the cell
2Reliability
If chemical effectors are used to control protein interactions, then the control can be achieved, but the system lacks spatial and temporal precision
Solution Approach 1:
The patent replaces chemical diffusion-based control with light-based optical control. Light can be focused to specific subcellular locations and turned on/off precisely in time, providing both spatial and temporal precision that chemical effectors cannot achieve due to their diffusive nature
Solution Approach 2:
The patent creates a dynamic control system where the phytochrome domain can rapidly switch between different conformational states (Pr and Pfr forms) in response to light signals. This dynamic responsiveness enables precise temporal control of protein interactions, allowing activation and deactivation on demand with millisecond precision
3Manufacturing precision
If light-regulated systems are used to control protein interactions, then spatial and temporal precision can be achieved, but the system lacks reversibility and specificity
Solution Approach 1:
The patent utilizes parameter changes in the phytochrome domain's light-absorption properties. The domain exists in two stable states: Pr (red-light absorbing) and Pfr (far-red-light absorbing). By changing the wavelength parameter of incident light, the system can reversibly switch between states, enabling specific and reversible control of protein interactions with high adaptability
Solution Approach 2:
The patent employs periodic alternating exposure to red and far-red light to achieve reversible control. Red light converts PhyB to the Pfr state that binds PIF, while far-red light converts it back to the Pr state that releases PIF. This periodic action enables repeated cycles of protein interaction activation and deactivation, providing full reversibility
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 provides precise, reversible, and detectable control over protein interactions within cells, allowing for rapid and localized modulation of biological processes, with the ability to recruit proteins to specific subcellular compartments and induce signaling cascades with high spatial and temporal resolution.
Implementation Method 1
The phytochrome domain can for example be initially converted to (i) the Pr state, thereby disallowing the interaction between the first protein construct and the second protein construct; or (ii) the Pfr state, thereby allowing the interaction between the first protein construct and the second protein construct
Data Source
AI summary
The invention provides methods, materials and systems of regulating association between proteins of interest using light. In an aspect, the invention takes advantage of the ability of phytochromes to change conformation upon exposure to appropriate light conditions, and to bind in a conformation-dependent manner to cognate proteins called phytochrome-interacting factors. The invention comprises a method of regulating interaction between a first protein of interest and second protein within a cell by light. Such a method optionally comprises providing in the cell (1) a first protein construct which comprises the first protein, a phytochrome domain (PHD), and (2) providing in the cell a second protein construct which comprises the second protein and a phytochrome domain-interacting peptide (PIP) that can bind selectively to the Pfr state, but not to the Pr state, of the phytochrome domain.