Source 1primary paper2013Biochemistry
Toolkit/EL222
EL222
Also known as: bacterial light-oxygen-voltage (LOV) photoreceptor EL222, blue light dependent DNA-binding protein EL222, light-activated EL222 transcription factor, LOV-HTH transcription factor, EL222
Taxonomy: Mechanism Branch / Architecture. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
EL222 is a blue light-activated LOV-HTH transcription factor from the marine bacterium Erythrobacter litoralis HTCC2594 that functions as a light-dependent DNA-binding protein for optical control of transcription. Its flavin mononucleotide chromophore photodynamics have been characterized in free solution and when embedded in EL222 variants.
Usefulness & Problems
Why this is useful
EL222 is useful as a genetically encoded light input module for regulating transcription with blue light through a native light-responsive DNA-binding protein. The cited literature also supports its value as a model LOV photoreceptor for dissecting FMN excited-state and adduct-state dynamics by time-resolved spectroscopy.
Source:
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
Source:
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
Problem solved
EL222 helps solve the problem of coupling a defined optical input to transcriptional regulation using a bacterial blue light-responsive transcription factor. The available evidence also addresses the mechanistic problem of resolving sub-millisecond FMN photochemistry in the EL222 LOV domain and its variants.
Source:
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
Published Workflows
Objective: Engineer a mammalian optogenetic gene-expression system with stronger light-induced expression suitable for basic research and bioproduction, including production of difficult-to-express complex antibodies.
Why it works: The paper states that insufficient expression and induction in existing mammalian optogenetic systems can be overcome by fusing the blue-light receptor EL222 to a stronger tandem activator module composed of VP64, p65, and Rta.
blue-light activated transcriptional control via EL222-based photoswitchingenhanced transcriptional activation through tandem VP64, p65, and Rta activator domainsfusion-protein designoptogenetic stimulationmammalian gene-expression testingapplication to recombinant antibody production
Stages
- 1.Photoswitch design(library_design)
This stage exists to redesign the optogenetic transcription factor so that mammalian cells can achieve stronger light-induced expression than current systems.
Selection: Fuse EL222 to stronger tandem transcriptional activator domains to overcome insufficient expression and induction in mammalian optogenetic systems.
- 2.Gene-expression performance testing(functional_characterization)
This stage tests whether the redesigned photoswitch actually overcomes the low induction and low expression limitations of prior mammalian optogenetic systems.
Selection: Measure blue-light induction of target gene expression and compare achieved expression to strong constitutive promoters.
- 3.Application to complex antibody production(confirmatory_validation)
This stage confirms that the strong light-inducible expression system is useful in a bioproduction-relevant application involving difficult-to-express proteins.
Selection: Use DEL-VPR to drive expression of complex monoclonal and bispecific antibodies and assess byproduct expression and yield of functional protein complexes.
Steps
- 1.Fuse EL222 to tandem VP64-p65-Rta activator domainsengineered photoswitch
Create a stronger blue-light inducible transcription factor for mammalian cells.
The abstract states this design was undertaken specifically to overcome insufficient expression levels and induction in current mammalian optogenetic gene-expression systems.
- 2.Test blue-light induction of target gene expressionphotoswitch under test
Determine whether DEL-VPR provides strong inducible expression in mammalian cells.
This performance test is needed after design to establish that the engineered construct overcomes the stated limitations of prior systems before applying it to complex protein production.
- 3.Apply DEL-VPR to light-induced expression of monoclonal and bispecific antibodiesexpression control system
Demonstrate utility of DEL-VPR for bioproduction-relevant expression of difficult protein complexes.
The application step follows expression testing because the paper uses antibody production as a downstream demonstration of practical value after establishing strong inducible expression.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Architecture: A composed arrangement of multiple parts that instantiates one or more mechanisms.
Mechanisms
blue light-induced dimerizationcysteinyl-flavin adduct formationHeterodimerizationlight-dependent dna bindinglov-domain fmn photochemistryTechniques
No technique tags yet.
Target processes
transcriptionInput: Light
Implementation Constraints
EL222 contains an FMN chromophore in its LOV photosensory module, so function depends on flavin-based photochemistry under blue light. The evidence explicitly references two EL222 variants and a set of 19 new variants, indicating that variant engineering has been applied, but the provided material does not specify construct architecture, promoter design, or delivery format.
The supplied evidence is strongest for photophysical characterization and only limited details are provided here on quantitative transcriptional performance, dynamic range, or host-range validation. Although 19 new EL222 variants are mentioned, the present evidence does not specify their functional properties or implementation outcomes.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2primary paper2016Nucleic Acids Research
Source 3review2022Trends in biotechnology
Source 4review2021FEMS Microbiology Reviews
Source 5primary paper2021Physical Chemistry Chemical Physics
Source 6review2021Advanced Biology
Source 7primary paper2023Biomolecules
Source 8primary paper2017Biochemistry
Ranked Claims
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Analysis of the low-frequency Raman region below 1000 cm^-1 provided evidence for additional dynamical events.
Importantly, we found evidence for additional dynamical events, particularly upon analysis of the low-frequency Raman region below 1000 cm-1.
low frequency region upper bound 1000 cm^-1
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
A broadband dual visTA/FSRS set-up spans approximately 200-2200 cm^-1 and supports delays from a few femtoseconds to several hundreds of microseconds after actinic pumping.
Here we report on a broadband (~200-2200 cm-1) dual transient visible absorption (visTA)/FSRS set-up that can accommodate time delays from a few femtoseconds to several hundreds of microseconds after illumination with an actinic pump.
time delay range a few femtoseconds to several hundreds of microsecondswavenumber range ~200-2200 cm^-1
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
We show that fs-to-sub-ms visTA/FSRS with a broad wavenumber range is a useful tool to characterize short-lived conformationally excited states in flavoproteins and potentially other light-responsive proteins.
Cph1 is discussed in connection with red/far-red-responsive extracellular matrix systems with reversibly tunable mechanical properties.
The anchor review cites the 2019 Adv Mater extracellular-matrix paper as using a cyanobacterial photoreceptor Cph1 for red/far-red mechanical tuning.
EL222 is discussed as a blue-light-responsive DNA-binding component for control of exopolysaccharide production and biofilm structure.
The anchor review and the Sinorhizobium meliloti primary paper support EL222 as a blue-light-responsive DNA-binding component for biofilm/exopolysaccharide control.
OptoAMP is discussed as a high-light-sensitivity blue-light system for production settings.
The anchor review explicitly discusses OptoAMP as a high-light-sensitivity blue-light system for production settings.
PULSE is discussed as a plant-usable light-switch system that combines EL222-based repression with a red-light-inducible activator architecture.
The anchor review explicitly names PULSE as a plant-usable light-switch system combining EL222-based repression with a red-light-inducible activator architecture.
The review explicitly supports Cph1, CRY2olig, EL222, LOV2, OptoAMP, and PULSE as named optogenetic components or tools within its scope.
Explicitly supported tool/component names found in these sources include Cph1, CRY2olig, EL222, LOV2, OptoAMP, and PULSE.
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
QM calculations were used to predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors.
QM calculations predict the energetics and infrared spectra of transient glutamine isomers in LOV photoreceptors
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
adduct formation rate variation 3.6 fold
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
structural change timescale micro- to submillisecondvariation magnitude orders of magnitude
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
structural change timescale micro- to submillisecondvariation magnitude orders of magnitude
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
structural change timescale micro- to submillisecondvariation magnitude orders of magnitude
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
structural change timescale micro- to submillisecondvariation magnitude orders of magnitude
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
structural change timescale micro- to submillisecondvariation magnitude orders of magnitude
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
structural change timescale micro- to submillisecondvariation magnitude orders of magnitude
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
The system was applied to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal.
We further apply the system, for the first time, to optogenetically synchronize two receiver cells performing different logic behaviors over time using blue light as a molecular clock signal
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
Gene expression level in the system can be precisely controlled by modulating blue-light pulse dosage or intensity.
by modulating the dosage of light pulses or intensity we could control the level of gene expression precisely
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The kinetics of light-inducible and repressible expression were quantitatively analyzed using a mathematical model.
the light-inducible and repressible expression kinetics were quantitatively analysed using a mathematical model
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
The light-inducible and light-repressible systems can function in parallel with high spatial precision in a single cell and can be switched stably between ON and OFF states by repetitive blue-light pulses.
both light-inducible and repressible system can function in parallel with high spatial precision in a single cell and can be switched stably between ON- and OFF-states by repetitive pulses of blue light
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
AQTrip oligomerizes in the absence of DNA and forms an EL222 dimer-DNA complex in the presence of DNA substrates.
Size-exclusion chromatography and light scattering indicate that AQTrip oligomerizes in the absence of DNA and selects for an EL222 dimer-DNA complex in the presence of DNA substrates
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
The AQTrip EL222 variant stabilizes the photoactivated state.
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
complex stoichiometry 2:1
Approval Evidence
9 sources17 linked approval claimsfirst-pass slug el222
We obtained 19 new variants of the LOV transcription factor El222
Source:
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
Source:
Explicitly supported tool/component names found in these sources include Cph1, CRY2olig, EL222, LOV2, OptoAMP, and PULSE.
Source:
The web research summary states that multiple discovered primary papers explicitly center EL222 as a bacterial blue-light transcription factor for optogenetic control.
Source:
EL222 from Erythrobacter litoralis
Source:
The web research summary identifies EL222 as an explicitly supported LOV-based optogenetic component relevant to the review's scope.
Source:
the LOV-HTH transcription factor, EL222
Source:
using the blue light dependent DNA-binding protein EL222
Source:
One such protein, the light-activated EL222 transcription factor, from the marine bacterium Erythrobacter litoralis HTCC2594
Source:
application capabilitysupports
Mutations yielding green-light-responsive El222 variants enabled orthogonal color-multiplexing using only LOV domains.
We demonstrate the utility of the latter mutations for orthogonal color-multiplexing with only LOV domains for the first time.
Source:
engineering resultsupports
The authors obtained 19 new El222 variants with stronger activity, lower leakiness, or green-light responsiveness in vivo.
We obtained 19 new variants of the LOV transcription factor El222 that were stronger, less leaky, or green light responsive in vivo .
Source:
agreement with prior worksupports
Observed lifetimes and intermediate states including singlet, triplet, and adduct agree with previous time-resolved infrared spectroscopy experiments.
The observed lifetimes and intermediate states (singlet, triplet, and adduct) are in agreement with previous time-resolved infrared spectroscopy experiments.
Source:
method applicationsupports
The extended time scale and wavenumber range allowed monitoring of the complete excited-state dynamics of FMN free in solution and FMN embedded in two EL222 variants.
The extended time scale and wavenumber range allowed us to monitor the complete excited-state dynamics of the biological chromophore flavin mononucleotide (FMN), both free in solution and embedded in two variants of the bacterial light-oxygen-voltage (LOV) photoreceptor EL222.
Source:
application summarysupports
EL222 is discussed as a blue-light-responsive DNA-binding component for control of exopolysaccharide production and biofilm structure.
The anchor review and the Sinorhizobium meliloti primary paper support EL222 as a blue-light-responsive DNA-binding component for biofilm/exopolysaccharide control.
Source:
component in scopesupports
The review explicitly supports Cph1, CRY2olig, EL222, LOV2, OptoAMP, and PULSE as named optogenetic components or tools within its scope.
Explicitly supported tool/component names found in these sources include Cph1, CRY2olig, EL222, LOV2, OptoAMP, and PULSE.
Source:
mechanistic inferencesupports
Calculated energies and rotational barriers for glutamine rotamers and tautomers allowed the authors to postulate the most energetically favoured glutamine orientation for each of EL222, AsLOV2, and RsLOV along the assumed reaction path.
Energies and rotational barriers were calculated for possible rotamers and tautomers of the critical glutamine side chain, which allowed us to postulate the most energetically favoured glutamine orientation for each LOV domain along the assumed reaction path.
Source:
mechanistic inferencesupports
Energetic and spectroscopic analyses converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and, more strongly, for AsLOV2, whereas RsLOV retains the initial glutamine configuration.
both the energetic and spectroscopic approaches converge in suggesting a facile glutamine flip at the adduct intermediate for EL222 and more so for AsLOV2, while for RsLOV the glutamine keeps its initial configuration
Source:
theory experiment agreementsupports
Constructed infrared difference spectra showed good agreement with experimental transient infrared spectra for EL222 and AsLOV2, permitting assignment of the majority of observed bands.
The good agreement between theory and experiment permitted the assignment of the majority of observed bands
Source:
mechanistic conservationsupports
Across AsLOV2, YtvA, EL222, and LovK, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved despite differences in tertiary structure.
Despite differences in tertiary structure, the overall pathway leading to cysteine adduct formation from the FMN triplet state is highly conserved, although there are slight variations in rate.
Source:
post adduct kinetic divergencesupports
After adduct formation, vibrational spectra and kinetics differ significantly among the full-length LOV photoreceptors and are directly linked to the specific output function of the LOV domain.
However, significant differences are observed in the vibrational spectra and kinetics after adduct formation, which are directly linked to the specific output function of the LOV domain.
Source:
rate variationsupports
The rate of adduct formation varies by only 3.6-fold among the studied LOV proteins.
While the rate of adduct formation varies by only 3.6-fold among the proteins
Source:
structural change timescalesupports
Large-scale structural changes in the full-length LOV photoreceptors occur over micro- to submillisecond time scales and vary by orders of magnitude depending on output function.
the subsequent large-scale structural changes in the full-length LOV photoreceptors occur over the micro- to submillisecond time scales and vary by orders of magnitude depending on the different output function of each LOV domain.
Source:
engineering resultsupports
The authors engineered a novel EL222-based bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly by blue light.
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
Source:
mechanismsupports
EL222 uses blue light to drive reorientation of LOV sensory and HTH effector domains, allowing photoactivation of gene transcription.
it harnesses blue light to drive the reorientation of light-oxygen-voltage (LOV) sensory and helix-turn-helix (HTH) effector domains to allow photoactivation of gene transcription in natural and artificial systems
Source:
mechanistic stepsupports
Blue light induces EL222 dimerization through LOV and HTH interfaces.
Combined, these novel approaches have validated a key mechanistic step, whereby blue light induces EL222 dimerization through LOV and HTH interfaces
Source:
stoichiometrysupports
The EL222-DNA complex has a 2:1 stoichiometry with a previously characterized DNA substrate.
NMR analyses of the EL222-DNA complex confirm a 2:1 stoichiometry in the presence of a previously characterized DNA substrate
Source:
Comparisons
Source-backed strengths
The tool is directly described as a blue light-dependent DNA-binding protein and light-activated transcription factor, supporting its use for optical transcription control. Spectroscopic studies monitored complete excited-state dynamics of FMN free in solution and in two EL222 variants, and the observed singlet, triplet, and adduct intermediates agreed with prior time-resolved infrared measurements. Additional low-frequency Raman features further indicate detectable dynamical events beyond the main intermediates.
Source:
we have engineered a novel bidirectional promoter system for Escherichia coli that can be induced or repressed rapidly and reversibly using the blue light dependent DNA-binding protein EL222
Source:
creating an EL222 variant harboring V41I, L52I, A79Q, and V121I point mutations (AQTrip) that stabilizes the photoactivated state
Ranked Citations
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