Source 1primary paper2020Figshare
Toolkit/SspB A58V iLID dimer variant
SspB A58V iLID dimer variant
Also known as: A58V variant
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The SspB A58V iLID dimer variant is a blue-light-inducible, multi-component protein interaction system in which a single A58V substitution in SspB tunes binding to iLID. It mediates light-gated heterodimerization and was reported to enable light-activated colocalization of transmembrane proteins in neurons.
Usefulness & Problems
Why this is useful
This variant is useful for optically controlling protein localization in contexts where effective protein concentrations are high. The reported affinity tuning reduced dark-state colocalization relative to a higher-affinity switch while preserving blue-light-induced recruitment in neurons.
Source:
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
Problem solved
It addresses the problem that earlier or higher-affinity iLID-SspB pairs can show excessive interaction in the dark when proteins are present at high effective concentrations. The reengineered interaction was reported to better control proteins present at effective concentrations of 5-100 μM.
Source:
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Techniques
No technique tags yet.
Target processes
localizationrecombinationInput: Light
Implementation Constraints
The system consists of iLID and an SspB partner carrying a single A58V point mutation. It is activated by blue light, and the cited engineering goal was improved control at high effective protein concentrations of 5-100 μM; no additional construct architecture, cofactor, or delivery details are provided in the supplied evidence.
The supplied evidence is limited to a single source and primarily supports neuronal transmembrane protein colocalization and affinity measurements. The evidence provided does not describe performance in other cell types, recombination applications, kinetic parameters for this specific variant, or implementation details beyond the A58V mutation.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
higher affinity switch range 0.8-47 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
binding affinity fold change 42binding affinity state 1 3 b1 2 bcMbinding affinity state 2 125 b1 40 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The iLID-SspB interaction was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
effective protein concentration range 5-100 bcM
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
The N414L point mutation in the LOV domain lengthened the reversion half-life of iLID.
with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
iLID contains a LOV domain that undergoes a conformational change upon blue-light activation and exposes the ssrA peptide motif that binds SspB.
iLID contains a light-oxygen-voltage (LOV) domain that undergoes a conformational change upon activation with blue light and exposes a peptide motif, ssrA, that binds to SspB
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
Approval Evidence
1 source3 linked approval claimsfirst-pass slug sspb-a58v-ilid-dimer-variant
The new variant of the dimer system contains a single SspB point mutation (A58V)
Source:
application performancesupports
The SspB A58V dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because more colocalization was seen in the dark.
allows for light-activated colocalization of transmembrane proteins in neurons, where a higher affinity switch (0.8-47 bcM) was less effective because more colocalization was seen in the dark
Source:
binding affinity changesupports
The SspB A58V dimer variant displays a 42-fold change in binding affinity upon blue-light activation, from 3 b1 2 bcM to 125 b1 40 bcM.
The new variant of the dimer system contains a single SspB point mutation (A58V), and displays a 42-fold change in binding affinity when activated with blue light (from 3 b1 2 bcM to 125 b1 40 bcM)
Source:
scope expansionsupports
The expanded suite of light-induced dimers increases the variety of cellular pathways that can be targeted with light.
This expanded suite of light induced dimers increases the variety of cellular pathways that can be targeted with light
Source:
Comparisons
Source-backed strengths
The A58V variant was reported to exhibit a 42-fold light-dependent affinity change, from 3 ± 2 μM to 125 ± 40 μM upon blue-light activation as stated in the source. In neuronal transmembrane protein colocalization experiments, it outperformed a higher-affinity switch because the higher-affinity system showed more colocalization in the dark.
Source:
we reengineer the interaction between the light inducible dimer, iLID, and its binding partner SspB, to better control proteins present at high effective concentrations (5-100 bcM)
Ranked Citations
- 1.