Source 1primary paper2020Scientific Reports
Toolkit/15N and 1H liquid-state high-resolution NMR
15N and 1H liquid-state high-resolution NMR
Also known as: 15N liquid-state high-resolution NMR, 1H liquid-state high-resolution NMR
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
15N and 1H liquid-state high-resolution NMR is an assay method used to detect light-induced photo-CIDNP signals in engineered Mr4511 flavoproteins. In the cited study, it reported nuclear hyperpolarization arising from a light-driven transient paramagnetic state in variants containing tryptophan at canonical or newly introduced positions.
Usefulness & Problems
Why this is useful
This assay is useful for directly observing photo-CIDNP and associated nuclear hyperpolarization in solution-phase flavoprotein systems under light activation. It enables evaluation of whether protein engineering, specifically tryptophan placement in Mr4511, produces detectable spin-chemistry outputs and supports mechanistic interpretation through magnetic-field dependence.
Problem solved
It addresses the problem of measuring light-driven nuclear hyperpolarization in engineered flavoproteins and determining whether introduced aromatic residues support photo-CIDNP-active states. It also helps probe whether the underlying transient paramagnetic state is influenced by anisotropic magnetic interactions and slow-motion behavior.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
anisotropic magnetic interactionsanisotropic magnetic interactionsnuclear hyperpolarizationnuclear hyperpolarizationphoto-cidnpphoto-cidnpslow-motion transient paramagnetic-state behaviorslow-motion transient paramagnetic-state behaviorTechniques
Functional AssayTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementmeasurement targets: 1H and 15Noperating role: sensor
The reported implementation involved 15N and 1H liquid-state high-resolution NMR measurements on engineered Mr4511 flavoproteins under light input. The assay was applied to variants containing tryptophan at canonical or novel positions, but the supplied evidence does not specify construct architecture, illumination parameters, isotope-labeling strategy, or expression and purification details.
The supplied evidence is limited to a single 2020 study in engineered Mr4511 flavoproteins, so broader generality across proteins, conditions, or assay formats is not established here. The evidence does not provide practical performance metrics such as sensitivity limits, throughput, sample requirements, or comparative benchmarking against other assays.
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
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 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:
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:
mechanistic interpretationsupports
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
Source:
Comparisons
Source-backed strengths
The method detected photo-CIDNP effects in engineered Mr4511 proteins with tryptophan inserted at both canonical and novel positions, demonstrating sensitivity to engineering-dependent changes in spin behavior. Magnetic-field-dependent measurements further provided mechanistic information consistent with anisotropic magnetic interactions and a slow-motion transient paramagnetic regime.
Ranked Citations
- 1.