Source 1primary paper2005Magnetic Resonance in Chemistry
Toolkit/site-directed spin labeling
site-directed spin labeling
Also known as: SDSL, spin labeling
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
In case the transfer process does not involve stable or transient paramagnetic species or states, site-directed spin labeling with suitable nitroxide radicals still allows EPR techniques to be used for studying structure and conformational dynamics of the proteins in action.
Usefulness & Problems
Why this is useful
Site-directed spin labeling introduces nitroxide radicals so EPR can probe protein structure and conformational dynamics even when the native system lacks paramagnetic states.; enabling EPR study of proteins that do not naturally contain stable or transient paramagnetic species; probing protein structure and conformational dynamics in action
Source:
Site-directed spin labeling introduces nitroxide radicals so EPR can probe protein structure and conformational dynamics even when the native system lacks paramagnetic states.
Source:
enabling EPR study of proteins that do not naturally contain stable or transient paramagnetic species
Source:
probing protein structure and conformational dynamics in action
Problem solved
It solves the problem of applying EPR to proteins whose transfer processes do not involve stable or transient paramagnetic species.; extends EPR applicability to otherwise EPR-inaccessible protein systems
Source:
It solves the problem of applying EPR to proteins whose transfer processes do not involve stable or transient paramagnetic species.
Source:
extends EPR applicability to otherwise EPR-inaccessible protein systems
Problem links
This assay is directly described as enabling EPR studies of protein structure and conformational dynamics in action, which is closely aligned with the gap around highly dynamic proteins. It could provide experimental distance constraints for fluctuating ensembles that are hard to capture with static structure prediction alone.
extends EPR applicability to otherwise EPR-inaccessible protein systems
LiteratureIt solves the problem of applying EPR to proteins whose transfer processes do not involve stable or transient paramagnetic species.
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It solves the problem of applying EPR to proteins whose transfer processes do not involve stable or transient paramagnetic species.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Mechanisms
Conformational Uncagingepr spectroscopic detection of conformational dynamicsnitroxide radical-based paramagnetic reportingsite-directed spin labelingTarget processes
No target processes tagged yet.
Input: Magnetic
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationoperating role: sensorswitch architecture: uncaging
The abstract states that suitable nitroxide radicals are required and that the method is used together with EPR instrumentation.; requires suitable nitroxide spin labels
Independent follow-up evidence is still limited. Validation breadth across biological contexts is still narrow. Independent reuse still looks limited, so the evidence base may be fragile. No canonical validation observations are stored yet, so context-specific performance remains under-specified.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
High-field EPR experiments with pH-sensitive nitroxide spin labels can be used to probe site-specific pK(a) values in protein systems.
to report on novel high-field EPR experiments for probing site-specific pK(a) values in protein systems by means of pH-sensitive nitroxide spin labels
Combining site-directed spin labeling with high-field/high-frequency EPR reveals unique information on proteins that is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
By combining site-directed spin labeling with high-field/high-frequency EPR, unique information on the proteins is revealed, which is complementary to that of X-ray crystallography, solid-state NMR, FRET, fast infrared and optical spectroscopic techniques.
High-field EPR can obtain detailed information on transient intermediates of proteins in biological action and can observe and characterize these intermediates while they remain in working states on biologically relevant timescales.
detailed information on the transient intermediates of the proteins in biological action is obtained. These intermediates can be observed and characterized while staying in their working states on biologically relevant timescales.
Site-directed spin labeling with suitable nitroxide radicals allows EPR to study protein structure and conformational dynamics even when the transfer process lacks stable or transient paramagnetic species or states.
In case the transfer process does not involve stable or transient paramagnetic species or states, site-directed spin labeling with suitable nitroxide radicals still allows EPR techniques to be used for studying structure and conformational dynamics of the proteins in action.
High-field EPR at 95 GHz/3.4 T and 360 GHz/12.9 T provides improved spectral and temporal resolution compared with conventional X-band EPR at 9.5 GHz/0.34 T.
Taking advantage of the improved spectral and temporal resolution of high-field EPR at 95 GHz/3.4 T and 360 GHz/12.9 T, as compared to conventional X-band EPR (9.5 GHz/0.34 T)
frequency 95 GHzfrequency 360 GHzfrequency 9.5 GHzmagnetic field 3.4 Tmagnetic field 12.9 Tmagnetic field 0.34 T
Approval Evidence
1 source2 linked approval claimsfirst-pass slug site-directed-spin-labeling
In case the transfer process does not involve stable or transient paramagnetic species or states, site-directed spin labeling with suitable nitroxide radicals still allows EPR techniques to be used for studying structure and conformational dynamics of the proteins in action.
Source:
method capabilitysupports
Combining site-directed spin labeling with high-field/high-frequency EPR reveals unique information on proteins that is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
By combining site-directed spin labeling with high-field/high-frequency EPR, unique information on the proteins is revealed, which is complementary to that of X-ray crystallography, solid-state NMR, FRET, fast infrared and optical spectroscopic techniques.
Source:
method capabilitysupports
Site-directed spin labeling with suitable nitroxide radicals allows EPR to study protein structure and conformational dynamics even when the transfer process lacks stable or transient paramagnetic species or states.
In case the transfer process does not involve stable or transient paramagnetic species or states, site-directed spin labeling with suitable nitroxide radicals still allows EPR techniques to be used for studying structure and conformational dynamics of the proteins in action.
Source:
Comparisons
Source-stated alternatives
The abstract says the resulting information is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
Source:
The abstract says the resulting information is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
Source-backed strengths
allows site-specific probing when combined with nitroxide radicals; can be combined with high-field/high-frequency EPR for additional information
Source:
allows site-specific probing when combined with nitroxide radicals
Source:
can be combined with high-field/high-frequency EPR for additional information
Compared with FRET
The abstract says the resulting information is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
Shared frame: source-stated alternative in extracted literature
Strengths here: allows site-specific probing when combined with nitroxide radicals; can be combined with high-field/high-frequency EPR for additional information.
Source:
The abstract says the resulting information is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
Compared with X-ray crystallography
The abstract says the resulting information is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
Shared frame: source-stated alternative in extracted literature
Strengths here: allows site-specific probing when combined with nitroxide radicals; can be combined with high-field/high-frequency EPR for additional information.
Source:
The abstract says the resulting information is complementary to X-ray crystallography, solid-state NMR, FRET, and fast infrared and optical spectroscopic techniques.
Ranked Citations
- 1.