Source 1primary paper2018
Toolkit/domain insertion permissibility
domain insertion permissibility
Also known as: domain insertion ‘permissibility’, permissibility
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
Domain insertion permissibility is an experimental engineering paradigm established in the human inward rectifier K+ channel Kir2.1 to identify engineerable allosteric sites. In this framework, sites permissive to insertion of regulatory domains can be converted into functional control points, including light-sensitive regulation when light-switchable domains are inserted.
Usefulness & Problems
Why this is useful
This method is useful for locating allosteric sites in Kir2.1 that can tolerate insertion of exogenous regulatory domains while remaining engineerable for control. The reported application shows that permissive or latent allosteric sites can be exploited to create light-responsive channel regulation, whereas nonpermissive sites do not support this outcome.
Source:
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Problem solved
It addresses the problem of identifying where regulatory domains can be inserted into an ion channel to create functional allosteric control. The source specifically frames it as a way to identify engineerable allosteric sites in human Kir2.1.
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete method used to build, optimize, or evolve an engineered system.
Techniques
Structural CharacterizationTarget processes
No target processes tagged yet.
Implementation Constraints
Implementation requires domain insertion into candidate sites within human Kir2.1 and functional assessment of the resulting constructs. The available evidence supports use of light-switchable inserted domains to test whether permissive or latent allosteric sites can confer light sensitivity, but it does not provide construct architecture, expression conditions, or cofactor requirements.
The evidence provided is centered on a single channel system, human Kir2.1, with only limited mention of equivalent sites in homologs. The literature also indicates that permissibility is context dependent and varies with the structural properties of the inserted domain, which may limit generalization across cargos and targets.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In support of this notion, inserting light-switchable domains into either existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Approval Evidence
1 source4 linked approval claimsfirst-pass slug domain-insertion-permissibility
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Source:
context dependencesupports
Many allosterically regulated sites in Kir2.1 or equivalent sites in homologs show differential permissibility that depends on the structural properties of the inserted domain.
Many allosterically regulated sites in Kir2.1 or sites equivalent to those regulated in homologs, such as G-protein-gated inward rectifier K + channels (GIRK), have differential permissibility; that is, for these sites permissibility depends on the structural properties of the inserted domain.
Source:
experimental paradigmsupports
Domain insertion permissibility is established as a new experimental paradigm to identify engineerable allosteric sites in human Kir2.1.
Here we use human Inward Rectifier K + Channel Kir2.1 to establish domain insertion ‘permissibility’ as a new experimental paradigm to identify engineerable allosteric sites.
Source:
mechanistic associationsupports
In Kir2.1, domain insertion permissibility is best explained by dynamic protein properties such as conformational flexibility.
We find that permissibility is best explained by dynamic protein properties, such as conformational flexibility.
Source:
proposed metricsupports
Differential permissibility is proposed as a metric of both existing and latent allostery in Kir2.1.
Our data and the well-established link between protein dynamics and allostery led us to propose that differential permissibility is a metric of both existing and latent allostery in Kir2.1.
Source:
Comparisons
Source-backed strengths
The paradigm is experimentally established in human Kir2.1 and links insertion permissibility to engineerable allostery. It further demonstrates functional specificity: inserting light-switchable domains into existing or latent allosteric sites, but not elsewhere, renders Kir2.1 activity sensitive to light.
Ranked Citations
- 1.