Source 1primary paper2004Biochemistry
Toolkit/Jalpha helix point mutations disrupting LOV-domain interaction
Jalpha helix point mutations disrupting LOV-domain interaction
Also known as: point mutations along the Jalpha helix
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Jalpha helix point mutations disrupting LOV-domain interaction are a phototropin construct pattern in which residues along the Jα helix are mutated to weaken LOV–Jα intramolecular association. In full-length phototropin, these mutations displace or unfold the Jα helix and produce constitutive kinase activation in the absence of illumination.
Usefulness & Problems
Why this is useful
This construct pattern is useful for probing how LOV-domain output is coupled to phototropin kinase regulation. By converting a light-regulated receptor into a constitutively active state, it enables mechanistic dissection of Jα-mediated signal transmission without requiring illumination.
Source:
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
Problem solved
It addresses the problem of identifying the structural event that links LOV-domain state changes to kinase activation in phototropin. The reported mutations specifically test whether weakening the LOV–Jα interaction is sufficient to activate signaling independently of light.
Problem links
Need conditional control of signaling activity
DerivedJalpha helix point mutations disrupting LOV-domain interaction are a phototropin construct pattern in which residues along the Jα helix are mutated to weaken LOV–Jα intramolecular association. In full-length phototropin, these mutations displace or unfold the Jα helix and produce constitutive kinase activation in the absence of illumination.
Need conditional protein clearance
DerivedJalpha helix point mutations disrupting LOV-domain interaction are a phototropin construct pattern in which residues along the Jα helix are mutated to weaken LOV–Jα intramolecular association. In full-length phototropin, these mutations displace or unfold the Jα helix and produce constitutive kinase activation in the absence of illumination.
Need precise spatiotemporal control with light input
DerivedJalpha helix point mutations disrupting LOV-domain interaction are a phototropin construct pattern in which residues along the Jα helix are mutated to weaken LOV–Jα intramolecular association. In full-length phototropin, these mutations displace or unfold the Jα helix and produce constitutive kinase activation in the absence of illumination.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A reusable architecture pattern for arranging parts into an engineered system.
Mechanisms
constitutive kinase activationDegradationdisruption of lov-jalpha intramolecular interactiondisruption of lov–jα intramolecular interactionjalpha helix displacementjalpha helix unfoldingjα-helix unfoldingTechniques
No technique tags yet.
Target processes
degradationsignalingInput: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: regulator
Implementation consists of point mutagenesis of residues along the phototropin Jα helix to disrupt its interaction with the LOV domain. The supplied evidence supports use in full-length phototropin constructs, but does not provide sequence-level design rules, expression systems, cofactors, or delivery details.
The evidence comes from a single cited study and is limited to phototropin, with no independent replication provided here. Specific mutated residues, quantitative activation levels, organismal context, and any effects on protein stability or degradation are not described in the supplied evidence.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
Unfolding of the Jalpha helix is the critical event regulating kinase signaling in phototropin proteins.
These results indicate that unfolding of the Jalpha helix is the critical event in regulation of kinase signaling for the phototropin proteins.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Approval Evidence
1 source2 linked approval claimsfirst-pass slug jalpha-helix-point-mutations-disrupting-lov-domain-interaction
we made a series of point mutations along the Jalpha helix to disrupt its interaction with the LOV domain
Source:
functional effectsupports
When introduced into full-length phototropin, the Jalpha helix point mutations cause constitutive kinase activation without illumination.
When placed into the full-length phototropin protein, these point mutations display constitutive kinase activation, without illumination of the sample.
Source:
mechanistic effectsupports
Several Jalpha helix point mutations displace the Jalpha helix from the LOV domain independently of illumination.
Using NMR spectroscopy and limited proteolysis, we demonstrate that several of these mutations displace the Jalpha helix from the LOV domain independently of illumination.
Source:
Comparisons
Source-backed strengths
The available evidence directly links Jα point mutagenesis to displacement of the Jα helix from the LOV domain and to constitutive activation of full-length phototropin kinase. The source also concludes that Jα helix unfolding is the critical regulatory event for kinase signaling, providing a clear mechanistic interpretation.
Compared with GFP-CRY2
Jalpha helix point mutations disrupting LOV-domain interaction and GFP-CRY2 address a similar problem space because they share degradation.
Shared frame: same top-level item type; shared target processes: degradation; shared mechanisms: degradation; same primary input modality: light
Compared with lyso-ArchT
Jalpha helix point mutations disrupting LOV-domain interaction and lyso-ArchT address a similar problem space because they share degradation.
Shared frame: same top-level item type; shared target processes: degradation; shared mechanisms: degradation; same primary input modality: light
Compared with PCB synthesis expression vector
Jalpha helix point mutations disrupting LOV-domain interaction and PCB synthesis expression vector address a similar problem space because they share degradation, signaling.
Shared frame: same top-level item type; shared target processes: degradation, signaling; shared mechanisms: degradation; same primary input modality: light
Strengths here: looks easier to implement in practice; may avoid an exogenous cofactor requirement.
Ranked Citations
- 1.