Source 1primary paper2016Biochemistry
Toolkit/iLID-SspB A58V variant
iLID-SspB A58V variant
Also known as: new variant of the dimer system, SspB A58V
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The iLID-SspB A58V variant is a blue-light-inducible heterodimerization system in which SspB carries a single A58V substitution. It was reengineered to tune iLID-SspB binding for improved light-controlled protein colocalization, including transmembrane protein localization in neurons.
Usefulness & Problems
Why this is useful
This variant is useful for optogenetic control of protein localization when lower dark-state association is needed at high effective intracellular concentrations. In neurons, it supported light-activated colocalization of transmembrane proteins more effectively than a higher-affinity switch because the higher-affinity system showed greater dark-state colocalization.
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 higher-affinity iLID-SspB switches can exhibit excessive dark-state colocalization, particularly in contexts with effective protein concentrations of 5-100 bcM. The A58V-tuned variant was reported as part of a reengineering effort to better control proteins under these high-concentration conditions.
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 variant is a multi-component construct requiring iLID and an SspB partner containing a single A58V point mutation. Its input modality is blue light, and the reported engineering goal was operation in settings with effective protein concentrations of 5-100 bcM; no additional construct architecture, cofactor, or delivery details are provided in the supplied evidence.
The supplied evidence is limited to one 2016 Biochemistry study and focuses on affinity tuning and neuronal transmembrane protein colocalization. No independent replication, detailed kinetic values for this specific variant, or broader validation across organisms, cell types, or downstream functional outputs are provided here.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Observations
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
successPrimary Cellsapplication demoneurons
light-activated colocalization
Inferred from claim c4 during normalization. The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization in the dark. Derived from claim c4. Quoted text: 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:
comparison switch affinity range(0.8-47)
Supporting Sources
Ranked Claims
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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
comparison switch affinity range 0.8-47 bcM
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The SspB A58V-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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 3 bcMbinding affinity 125 bcMfold change in binding affinity 42
The iLID-SspB system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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 system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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 system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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 system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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 system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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 system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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 system was reengineered to better control proteins present at high effective concentrations of 5-100 bcM.
Here, 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
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID.
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID.
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID.
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID.
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID.
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, with a point mutation in the LOV domain (N414L), we lengthened the reversion half-life of iLID.
A point mutation in the LOV domain, N414L, lengthened the reversion half-life of iLID.
Additionally, 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 ilid-sspb-a58v-variant
The new variant of the dimer system contains a single SspB point mutation (A58V)
Source:
application performancesupports
The SspB A58V-containing iLID dimer variant allows light-activated colocalization of transmembrane proteins in neurons, whereas a higher-affinity switch was less effective because it showed more colocalization 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-containing iLID dimer variant displays a 42-fold light-dependent change in binding affinity, from 125 bcM in one state to 3 bcM in the activated blue-light state.
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-containing variant showed a 42-fold light-dependent affinity change, from 125 bcM in one state to 3 bcM in the blue-light-activated state. It was specifically reported to enable light-activated colocalization of transmembrane proteins in neurons with reduced dark-state colocalization relative to a higher-affinity switch.
Source:
Here, 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.