Source 1primary paper2020ACS Chemical Biology
Toolkit/Zdk2-AsLOV2 optogenetic construct
Zdk2-AsLOV2 optogenetic construct
Also known as: Zdk2-AsLOV2
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Zdk2-AsLOV2 is an optogenetic protein-domain construct built around the Avena sativa LOV2 photosensory module. The supplied evidence supports blue-light activation through canonical AsLOV2 photochemistry, including cysteinyl-FMN adduct formation and Jα-helix unfolding, but does not describe the specific functional role contributed by the Zdk2 fusion in cells.
Usefulness & Problems
Why this is useful
This construct is useful as a blue-light-responsive protein module because the evidence supports a defined conformational uncaging mechanism in AsLOV2. The available literature clarifies how FMN excitation is coupled to structural rearrangements, which is valuable for mechanistic interpretation of AsLOV2-based optogenetic designs.
Problem solved
It helps address the problem of converting blue-light input into a predictable protein conformational change via the AsLOV2 domain. The supplied evidence does not specify which downstream cellular process the Zdk2-AsLOV2 construct was designed to control.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Mechanisms
blue-light-induced cysteinyl-fmn adduct formationconformational uncagingConformational Uncagingjα-helix unfoldingTechniques
Structural CharacterizationTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
The construct is based on the AsLOV2 light-sensing module and therefore depends on FMN-associated LOV photochemistry under blue-light illumination. The evidence specifically highlights the dark-state N414-Q513 hydrogen-bonding arrangement and light-triggered Jα-helix uncaging, but provides no construct architecture, linker, expression system, or delivery details for Zdk2-AsLOV2.
The evidence is mechanistic and centered on the AsLOV2 photosensor rather than on the full Zdk2-AsLOV2 construct's cellular performance. No data are provided here on expression, dynamic range, reversibility in cells, kinetics of the fusion construct, or validated biological outputs.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
In LOV2, blue light activation leads to formation of a Cys-FMN adduct, rotation of Q513, and unfolding of the Jα helix.
In the C-terminal light, oxygen, voltage (LOV) domain of plant phototropins (LOV2), blue light activation leads to formation of an adduct between a conserved Cys residue and the embedded FMN chromophore, rotation of a conserved Gln (Q513), and unfolding of a helix (Jα-helix)
In LOV2, blue light activation leads to formation of a Cys-FMN adduct, rotation of Q513, and unfolding of the Jα helix.
In the C-terminal light-oxygen-voltage (LOV) domain of plant phototropins (LOV2), blue light activation leads to formation of an adduct between a conserved Cys residue and the embedded FMN chromophore, rotation of a conserved Gln (Q513), and unfolding of a helix (Jα-helix)
In the dark state of AsLOV2, the side chain of N414 is hydrogen bonded to the backbone N-H of Q513.
In the dark state, the side chain of N414 is hydrogen bonded to the backbone N-H of Q513.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
conformational change timescale microsecondexcitation timescale sub-ps
Simulations predict that after Cys adduct formation, Q513 undergoes a lever-like motion that disrupts the N414-Q513 backbone interaction and forms a transient side-chain hydrogen bond between Q513 and N414.
The simulations predict a lever-like motion of Q513 after Cys adduct formation resulting in loss of the interaction between the side chain of N414 and the backbone C=O of Q513, and formation of a transient hydrogen bond between the Q513 and N414 side chains.
molecular dynamics simulation duration 7 μcs
Simulations predict that after Cys adduct formation, Q513 undergoes a lever-like motion that disrupts the N414-Q513 backbone interaction and forms a transient side-chain hydrogen bond between Q513 and N414.
The simulations predict a lever-like motion of Q513 after Cys adduct formation resulting in a loss of the interaction between the side chain of N414 and the backbone C═O of Q513, and formation of a transient hydrogen bond between the Q513 and N414 side chains.
In the dark state of AsLOV2, the side chain of N414 is hydrogen bonded to the backbone N-H of Q513.
In the dark state, the side chain of N414 is hydrogen bonded to the backbone N-H of Q513.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug zdk2-aslov2-optogenetic-construct
the cellular function of the Zdk2-AsLOV2 optogenetic construct
Source:
mechanismsupports
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Source:
mechanismsupports
Q513 and N414 are critical mediators of protein structural dynamics linking ultrafast FMN excitation to microsecond conformational changes that result in photoreceptor activation and biological function.
Through this multifaceted approach, we show that Q513 and N414 are critical mediators of protein structural dynamics, linking the ultrafast (sub-ps) excitation of the FMN chromophore to the microsecond conformational changes that result in photoreceptor activation and biological function.
Source:
structure function linksupports
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Source:
structure function relationshipsupports
Site-directed mutagenesis supports a direct link between Jα helix unfolding dynamics and cellular function of the Zdk2-AsLOV2 optogenetic construct.
The central role of N414 in signal transduction was evaluated by site-directed mutagenesis supporting a direct link between Jα helix unfolding dynamics and the cellular function of the Zdk2-AsLOV2 optogenetic construct.
Source:
Comparisons
Source-backed strengths
The mechanism is supported by a specific residue-level model in which blue light induces a Cys-FMN adduct, Q513 rotation, disruption of the N414-Q513 interaction, and Jα-helix unfolding. The cited study links ultrafast FMN excitation to microsecond conformational changes and identifies Q513 and N414 as critical mediators of activation dynamics.
Ranked Citations
- 1.