Source 1primary paper2017The Journal of Physical Chemistry B
Toolkit/time-resolved vibrational spectroscopy
time-resolved vibrational spectroscopy
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Time-resolved vibrational spectroscopy coupled with isotope labeling is an assay method used to resolve light-triggered structural dynamics in the Avena sativa LOV2 (AsLOV2) photosensory domain. In the cited study, it mapped structural evolution from 100 fs to 1 ms after optical excitation and supported a sequential allosteric model linking the flavin pocket to Jα-helix unfolding.
Usefulness & Problems
Why this is useful
This method is useful for assigning the temporal order of structural changes during photoactivation in a light-sensitive protein domain. In AsLOV2, it enabled observation of signal propagation from the flavin-binding pocket to the beta-sheet and then alpha-helix regions over femtosecond-to-millisecond timescales.
Problem solved
It addresses the problem of experimentally resolving ultrafast-to-slower structural intermediates in light-induced allostery within AsLOV2. The cited work used it to define a sequential pathway from initial flavin-pocket events to formation of the signaling state associated with Jα-helix unfolding.
Problem links
Need precise spatiotemporal control with light input
DerivedTime-resolved vibrational spectroscopy coupled with isotope labeling is an assay method used to map light-triggered structural dynamics in the Avena sativa LOV2 (AsLOV2) domain. In the cited study, it resolved structural evolution from 100 fs to 1 ms after optical excitation and supported a sequential allosteric model from the flavin pocket to Jα-helix unfolding.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
light-induced allosteric switchinglight-induced allosteric switchingsignal propagation through sequential structural rearrangementsignal propagation through sequential structural rearrangementTechniques
Directed EvolutionFunctional AssayFunctional AssayStructural CharacterizationStructural CharacterizationTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: sensorswitch architecture: uncaging
The method was applied under optical excitation and incorporated isotope labeling to aid assignment of structural evolution in AsLOV2. The available evidence does not provide further practical details on sample preparation, labeling scheme, instrument configuration, or expression system.
The supplied evidence is limited to one 2017 study on the AsLOV2 domain, so generality to other proteins or assay contexts is not established here. The evidence provided does not specify instrumentation details, spectral bands, throughput, or comparative performance against other time-resolved structural methods.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Point mutagenesis testing supports a key mediating role for Q513 in the AsLOV2 allosteric model.
This model is tested through point mutagenesis, elucidating in particular the key mediating role played by Q513.
Point mutagenesis testing supports a key mediating role for Q513 in the AsLOV2 allosteric model.
This model is tested through point mutagenesis, elucidating in particular the key mediating role played by Q513.
Point mutagenesis testing supports a key mediating role for Q513 in the AsLOV2 allosteric model.
This model is tested through point mutagenesis, elucidating in particular the key mediating role played by Q513.
Point mutagenesis testing supports a key mediating role for Q513 in the AsLOV2 allosteric model.
This model is tested through point mutagenesis, elucidating in particular the key mediating role played by Q513.
Point mutagenesis testing supports a key mediating role for Q513 in the AsLOV2 allosteric model.
This model is tested through point mutagenesis, elucidating in particular the key mediating role played by Q513.
In AsLOV2, the earliest light-induced events occur in the flavin binding pocket, followed by structural changes in the beta-sheet and then alpha-helix regions, culminating in Jb1-helix unfolding that yields the signaling state.
The earliest events occur in the flavin binding pocket, where a subpicosecond perturbation of the protein matrix occurs. In this perturbed environment, the previously characterized reaction between triplet state isoalloxazine and an adjacent cysteine leads to formation of the adduct state; this step is shown to exhibit dispersive kinetics. This reaction promotes coupling of the optical excitation to successive time-dependent structural changes, initially in the b2-sheet and then b1-helix regions of the AsLOV2 domain, which ultimately gives rise to Jb1-helix unfolding, yielding the signaling state.
In AsLOV2, the earliest light-induced events occur in the flavin binding pocket, followed by structural changes in the beta-sheet and then alpha-helix regions, culminating in Jb1-helix unfolding that yields the signaling state.
The earliest events occur in the flavin binding pocket, where a subpicosecond perturbation of the protein matrix occurs. In this perturbed environment, the previously characterized reaction between triplet state isoalloxazine and an adjacent cysteine leads to formation of the adduct state; this step is shown to exhibit dispersive kinetics. This reaction promotes coupling of the optical excitation to successive time-dependent structural changes, initially in the b2-sheet and then b1-helix regions of the AsLOV2 domain, which ultimately gives rise to Jb1-helix unfolding, yielding the signaling state.
In AsLOV2, the earliest light-induced events occur in the flavin binding pocket, followed by structural changes in the beta-sheet and then alpha-helix regions, culminating in Jb1-helix unfolding that yields the signaling state.
The earliest events occur in the flavin binding pocket, where a subpicosecond perturbation of the protein matrix occurs. In this perturbed environment, the previously characterized reaction between triplet state isoalloxazine and an adjacent cysteine leads to formation of the adduct state; this step is shown to exhibit dispersive kinetics. This reaction promotes coupling of the optical excitation to successive time-dependent structural changes, initially in the b2-sheet and then b1-helix regions of the AsLOV2 domain, which ultimately gives rise to Jb1-helix unfolding, yielding the signaling state.
In AsLOV2, the earliest light-induced events occur in the flavin binding pocket, followed by structural changes in the beta-sheet and then alpha-helix regions, culminating in Jb1-helix unfolding that yields the signaling state.
The earliest events occur in the flavin binding pocket, where a subpicosecond perturbation of the protein matrix occurs. In this perturbed environment, the previously characterized reaction between triplet state isoalloxazine and an adjacent cysteine leads to formation of the adduct state; this step is shown to exhibit dispersive kinetics. This reaction promotes coupling of the optical excitation to successive time-dependent structural changes, initially in the b2-sheet and then b1-helix regions of the AsLOV2 domain, which ultimately gives rise to Jb1-helix unfolding, yielding the signaling state.
In AsLOV2, the earliest light-induced events occur in the flavin binding pocket, followed by structural changes in the beta-sheet and then alpha-helix regions, culminating in Jb1-helix unfolding that yields the signaling state.
The earliest events occur in the flavin binding pocket, where a subpicosecond perturbation of the protein matrix occurs. In this perturbed environment, the previously characterized reaction between triplet state isoalloxazine and an adjacent cysteine leads to formation of the adduct state; this step is shown to exhibit dispersive kinetics. This reaction promotes coupling of the optical excitation to successive time-dependent structural changes, initially in the b2-sheet and then b1-helix regions of the AsLOV2 domain, which ultimately gives rise to Jb1-helix unfolding, yielding the signaling state.
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
time window end 1 mstime window start 100 fs
Approval Evidence
1 source1 linked approval claimfirst-pass slug time-resolved-vibrational-spectroscopy
Using time-resolved vibrational spectroscopy coupled with isotope labeling
Source:
structural dynamics mappingsupports
Time-resolved vibrational spectroscopy coupled with isotope labeling mapped structural evolution of AsLOV2 between 100 fs and 1 ms after optical excitation.
we have mapped the structural evolution of the LOV2 domain of the flavin binding phototropin Avena sativa (AsLOV2) over 10 decades of time, reporting structural dynamics between 100 fs and 1 ms after optical excitation
Source:
Comparisons
Source-backed strengths
A key strength is the broad temporal coverage, spanning 100 fs to 1 ms after optical excitation in a single mechanistic framework. Coupling vibrational spectroscopy with isotope labeling provided structural mapping that was sufficient to support a residue-level allosteric model in which Q513 has a mediating role.
Compared with native green gel system
time-resolved vibrational spectroscopy and native green gel system address a similar problem space.
Shared frame: same top-level item type; same primary input modality: light
Compared with open-source microplate reader
time-resolved vibrational spectroscopy and open-source microplate reader address a similar problem space.
Shared frame: same top-level item type; same primary input modality: light
Compared with plant transcriptome profiling
time-resolved vibrational spectroscopy and plant transcriptome profiling address a similar problem space.
Shared frame: same top-level item type; same primary input modality: light
Ranked Citations
- 1.