Source 1primary paper2020Scientific Reports
Toolkit/Mr4511 LOV domain
Mr4511 LOV domain
Also known as: a LOV domain named 4511 from Methylobacterium radiotolerans, Mr4511
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Mr4511 is a light-responsive LOV domain from Methylobacterium radiotolerans that has been used as a protein scaffold for engineered flavoprotein spin machines. In designed variants, tryptophan insertion at canonical or novel positions enabled illumination-dependent nuclear hyperpolarization detectable by 15N and 1H liquid-state high-resolution NMR.
Usefulness & Problems
Why this is useful
This domain is useful as a genetically encoded flavoprotein scaffold for creating light-driven systems that generate nuclear hyperpolarization. The reported function provides a route to optically induced photo-CIDNP signals observable by high-resolution liquid-state NMR.
Problem solved
Mr4511 helps address the challenge of engineering protein-based, light-activated spin systems that can pump nuclear hyperpolarization. The reported designs specifically solve this by introducing tryptophan residues into the LOV-domain scaffold to produce detectable photo-CIDNP effects.
Published Workflows
Objective: Design LOV-domain flavoproteins that act as light-driven spin machines producing nuclear hyperpolarization upon illumination.
Why it works: The abstract states that illumination drives FMN to abstract an electron from tryptophan, forming a transient spin-correlated radical pair that generates the photo-CIDNP effect.
FMN-mediated electron abstraction from tryptophantransient spin-correlated radical pair formationheuristic biomimetic designliquid-state high-resolution NMR readout
Stages
- 1.Biomimetic LOV-domain design(library_design)
The abstract describes a heuristic biomimetic design strategy using LOV domains and tryptophan engineering to create molecular spin machines.
Selection: Choose LOV-domain flavoprotein scaffolds and engineer tryptophan placement to enable photo-CIDNP-generating radical-pair chemistry.
- 2.NMR-based observation of photo-CIDNP(confirmatory_validation)
The abstract uses NMR observation as the evidence that the engineered designs produce photo-CIDNP and to infer magnetic interaction features.
Selection: Observe photo-CIDNP effects by 15N and 1H liquid-state high-resolution NMR.
Steps
- 1.Select LOV-domain scaffolds for spin-machine designengineered protein scaffolds
Use LOV-domain flavoproteins as the basis for designed molecular spin machines.
Scaffold choice precedes residue-level engineering because the design is framed around specific LOV-domain proteins.
- 2.Insert tryptophan at canonical and novel positions in Mr4511engineered protein scaffold
Create the tryptophan-containing design needed for the reported photo-CIDNP effect in Mr4511.
The abstract identifies tryptophan insertion as the engineering change that enables the observed effect in a scaffold whose wild-type form lacks the otherwise conserved tryptophan.
- 3.Measure engineered variants by 15N and 1H liquid-state high-resolution NMRengineered construct and assay readout
Observe whether the engineered variants yield photo-CIDNP effects and assess magnetic-field dependence.
NMR is used after design to confirm that the engineered proteins produce the intended hyperpolarization behavior.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Mechanisms
anisotropic magnetic interactions in a transient paramagnetic statelight-driven nuclear hyperpolarizationphoto-cidnpTechniques
Computational DesignTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
The reported implementations used designed LOV-domain flavoproteins based on the Mr4511 scaffold and required tryptophan insertion to obtain the photo-CIDNP effect. Function was assayed under illumination by detecting nuclear hyperpolarization with 15N and 1H liquid-state high-resolution NMR; no further construct, cofactor, or expression details are provided in the supplied evidence.
The available evidence is limited to a single 2020 study and focuses on engineered variants rather than the native Mr4511 domain alone. The supplied evidence does not report quantitative performance metrics, cellular validation, or applications beyond NMR-detectable photo-CIDNP.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Insertion of tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects detectable by 15N and 1H liquid-state high-resolution NMR.
Insertion of the tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects observed by 15N and 1H liquid-state high-resolution NMR
Insertion of tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects detectable by 15N and 1H liquid-state high-resolution NMR.
Insertion of the tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects observed by 15N and 1H liquid-state high-resolution NMR
Insertion of tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects detectable by 15N and 1H liquid-state high-resolution NMR.
Insertion of the tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects observed by 15N and 1H liquid-state high-resolution NMR
Insertion of tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects detectable by 15N and 1H liquid-state high-resolution NMR.
Insertion of the tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects observed by 15N and 1H liquid-state high-resolution NMR
Insertion of tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects detectable by 15N and 1H liquid-state high-resolution NMR.
Insertion of the tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects observed by 15N and 1H liquid-state high-resolution NMR
Designed LOV-domain flavoproteins can produce nuclear hyperpolarization upon illumination.
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination
Designed LOV-domain flavoproteins can produce nuclear hyperpolarization upon illumination.
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination
Designed LOV-domain flavoproteins can produce nuclear hyperpolarization upon illumination.
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination
Designed LOV-domain flavoproteins can produce nuclear hyperpolarization upon illumination.
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination
Designed LOV-domain flavoproteins can produce nuclear hyperpolarization upon illumination.
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination
The magnetic-field dependence of the observed photo-CIDNP effects indicates involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state.
with a characteristic magnetic-field dependence indicating an involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state
The magnetic-field dependence of the observed photo-CIDNP effects indicates involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state.
with a characteristic magnetic-field dependence indicating an involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state
The magnetic-field dependence of the observed photo-CIDNP effects indicates involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state.
with a characteristic magnetic-field dependence indicating an involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state
The magnetic-field dependence of the observed photo-CIDNP effects indicates involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state.
with a characteristic magnetic-field dependence indicating an involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state
The magnetic-field dependence of the observed photo-CIDNP effects indicates involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state.
with a characteristic magnetic-field dependence indicating an involvement of anisotropic magnetic interactions and a slow-motion regime in the transient paramagnetic state
Approval Evidence
1 source2 linked approval claimsfirst-pass slug mr4511-lov-domain
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination: ... a LOV domain named 4511 from Methylobacterium radiotolerans (Mr4511) which lacks an otherwise conserved tryptophan in its wild-type form.
Source:
engineering resultsupports
Insertion of tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects detectable by 15N and 1H liquid-state high-resolution NMR.
Insertion of the tryptophan at canonical and novel positions in Mr4511 yields photo-CIDNP effects observed by 15N and 1H liquid-state high-resolution NMR
Source:
functional capabilitysupports
Designed LOV-domain flavoproteins can produce nuclear hyperpolarization upon illumination.
Here, we report on designed molecular spin-machines producing nuclear hyperpolarization upon illumination
Source:
Comparisons
Source-backed strengths
Engineered Mr4511 variants produced illumination-dependent nuclear hyperpolarization with detection in both 15N and 1H liquid-state high-resolution NMR. The source specifically reports successful function from tryptophan insertion at both canonical and novel positions, indicating some positional flexibility in design.
Ranked Citations
- 1.