Source 1primary paper2023Angewandte Chemie International Edition
Toolkit/TiGGER
TiGGER
Also known as: time-resolved Gd-Gd electron paramagnetic resonance
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
TiGGER is a 240 GHz time-resolved Gd-Gd electron paramagnetic resonance assay for tracking inter-residue distances during a protein mechanical cycle in the solution state. It was demonstrated on the light-responsive AsLOV2 domain to resolve time-dependent structural separation and relaxation after photoactivation.
Usefulness & Problems
Why this is useful
TiGGER is useful for monitoring triggered protein conformational dynamics in solution through time-resolved distance measurements between Gd-labeled sites. The source literature positions it as a method that can complement existing approaches for studying functional protein dynamics.
Source:
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein's mechanical cycle in the solution state.
Source:
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein’s mechanical cycle in the solution state.
Problem solved
TiGGER addresses the problem of following inter-residue distance changes during a protein mechanical cycle in real time in solution. In the reported application, it enabled measurement of light-triggered long-range structural motion in AsLOV2, including separation and return to equilibrium.
Problem links
TiGGER provides time-resolved structural information on protein conformational cycles, which could contribute one high-resolution biomolecular layer of state information. However, the evidence is focused on inter-residue distances in a protein mechanical cycle rather than general multimodal cellular-state capture.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
gd-gd electron paramagnetic resonance distance measurementgd-gd electron paramagnetic resonance distance measurementlight-triggered protein conformational dynamicslight-triggered protein conformational dynamicstime-resolved tracking of inter-residue distance changestime-resolved tracking of inter-residue distance changesTechniques
Functional AssayTarget processes
No target processes tagged yet.
Input: Magnetic
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensor
The assay is a high-field 240 GHz time-resolved Gd-Gd electron paramagnetic resonance measurement performed in the solution state. The evidence supports use with light-triggered protein dynamics in AsLOV2, but it does not provide construct design details, Gd-label chemistry, or instrumentation setup beyond the operating frequency.
The supplied evidence describes application only to AsLOV2 and does not report broader benchmarking across proteins or assay formats. Practical performance parameters such as distance range, sensitivity, labeling requirements, and throughput are not provided in the evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successCell-freeapplication demo
Inferred from claim c3 during normalization. Upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s. Derived from claim c3. Section: abstract. Quoted text: Using TiGGER, we determined that upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s.
Source:
relaxation time constant60 sseparation time1 s(<)
successCell-freeapplication demo
Inferred from claim c4 during normalization. The Q513A variant of AsLOV2 shows slowed light-activated long-range mechanical motion, correlated with similarly slowed relaxation of the optically excited chromophore. Derived from claim c4. Section: abstract. Quoted text: TiGGER revealed that the light-activated long-range mechanical motion is slowed in the Q513A variant of AsLOV2 and is correlated to the similarly slowed relaxation of the optically excited chromophore as described in recent literature.
Source:
Supporting Sources
Source 2primary paper2022
Ranked Claims
TiGGER is a 240 GHz time-resolved Gd-Gd electron paramagnetic resonance method for tracking inter-residue distances during a protein mechanical cycle in solution.
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein's mechanical cycle in the solution state.
Section: abstract
frequency 240 GHz
Upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s.
Using TiGGER, we determined that upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s.
Section: abstract
relaxation time constant 60 sseparation time 1 s
TiGGER has the potential to complement existing methods for studying triggered functional dynamics in proteins.
TiGGER has the potential to valuably complement existing methods for the study of triggered functional dynamics in proteins.
Section: abstract
The Q513A variant of AsLOV2 shows slowed light-activated long-range mechanical motion, correlated with similarly slowed relaxation of the optically excited chromophore.
TiGGER revealed that the light-activated long-range mechanical motion is slowed in the Q513A variant of AsLOV2 and is correlated to the similarly slowed relaxation of the optically excited chromophore as described in recent literature.
Section: abstract
TiGGER at 240 GHz can track inter-residue distances during a protein mechanical cycle in solution state.
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein’s mechanical cycle in the solution state.
frequency 240 GHz
TiGGER has the potential to complement existing methods for studying triggered functional dynamics in proteins.
TiGGER has the potential to valuably complement existing methods for the study of triggered functional dynamics in proteins.
Approval Evidence
2 sources6 linked approval claimsfirst-pass slug tigger
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein's mechanical cycle in the solution state.
Source:
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein’s mechanical cycle in the solution state.
Source:
capabilitysupports
TiGGER is a 240 GHz time-resolved Gd-Gd electron paramagnetic resonance method for tracking inter-residue distances during a protein mechanical cycle in solution.
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein's mechanical cycle in the solution state.
Source:
observed dynamicssupports
Upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s.
Using TiGGER, we determined that upon light activation, the C-terminus and N-terminus of AsLOV2 separate in less than 1 s and relax back to equilibrium with a time constant of approximately 60 s.
Source:
positioningsupports
TiGGER has the potential to complement existing methods for studying triggered functional dynamics in proteins.
TiGGER has the potential to valuably complement existing methods for the study of triggered functional dynamics in proteins.
Source:
variant effectsupports
The Q513A variant of AsLOV2 shows slowed light-activated long-range mechanical motion, correlated with similarly slowed relaxation of the optically excited chromophore.
TiGGER revealed that the light-activated long-range mechanical motion is slowed in the Q513A variant of AsLOV2 and is correlated to the similarly slowed relaxation of the optically excited chromophore as described in recent literature.
Source:
method capabilitysupports
TiGGER at 240 GHz can track inter-residue distances during a protein mechanical cycle in solution state.
We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein’s mechanical cycle in the solution state.
Source:
positioning statementsupports
TiGGER has the potential to complement existing methods for studying triggered functional dynamics in proteins.
TiGGER has the potential to valuably complement existing methods for the study of triggered functional dynamics in proteins.
Source:
Comparisons
Source-backed strengths
The method directly tracks inter-residue distances during dynamic protein function in solution at 240 GHz. In AsLOV2, it resolved that the N-terminus and C-terminus separate in less than 1 s after light activation and relax with an approximately 60 s time constant, and it detected slowed long-range motion in the Q513A variant.
Compared with native green gel system
TiGGER and native green gel system address a similar problem space.
Shared frame: same top-level item type
Strengths here: appears more independently replicated; looks easier to implement in practice.
Compared with Nitrogen Vacancy diamond centers
TiGGER and Nitrogen Vacancy diamond centers address a similar problem space.
Shared frame: same top-level item type; same primary input modality: magnetic
Strengths here: appears more independently replicated; looks easier to implement in practice.
Compared with time-resolved Gd-Gd electron paramagnetic resonance
TiGGER and time-resolved Gd-Gd electron paramagnetic resonance address a similar problem space.
Shared frame: same top-level item type; shared mechanisms: time-resolved tracking of inter-residue distance changes; same primary input modality: magnetic
Strengths here: appears more independently replicated; looks easier to implement in practice.
Ranked Citations
- 1.
Derived from 4 linked claims and 2 validation observations. Example evidence: We present time-resolved Gd-Gd electron paramagnetic resonance (TiGGER) at 240 GHz for tracking inter-residue distances during a protein's mechanical cycle in the solution state.
- 2.