Conformational Coupling Research Articles

Dissected antiporter modules establish minimal proton-conduction elements of the respiratory complex I

The respiratory Complex I is a highly intricate redox-driven proton pump that powers oxidative phosphorylation across all domains of life. Yet, despite major efforts in recent decades, its long-range energy transduction principles remain highly debated. We create here minimal proton-conducting membrane modules by engineering and dissecting the key elements of the bacterial Complex I. By combining biophysical, biochemical, and computational experiments, we show that the isolated antiporter-like modules of Complex I comprise all functional elements required for conducting protons across proteoliposome membranes. We find that the rate of proton conduction is controlled by conformational changes of buried ion-pairs that modulate the reaction barriers by electric field effects. The proton conduction is also modulated by bulky residues along the proton channels that are key for establishing a tightly coupled proton pumping machinery in Complex I. Our findings provide direct experimental evidence that the individual antiporter modules are responsible for the proton transport activity of Complex I. On a general level, our findings highlight electrostatic and conformational coupling mechanisms in the modular energy-transduction machinery of Complex I with distinct similarities to other enzymes.

Testing scale-invariant inflation against cosmological data

There is solid theoretical and observational motivation behind the idea of scale-invariance as a fundamental symmetry of Nature. We consider a recently proposed classically scale-invariant inflationary model, quadratic in curvature and featuring a scalar field non-minimally coupled to gravity. We go beyond earlier analytical studies, which showed that the model predicts inflationary observables in qualitative agreement with data, by solving the full two-field dynamics of the system — this allows us to corroborate previous analytical findings and set robust constraints on the model's parameters using the latest Cosmic Microwave Background (CMB) data from Planck and BICEP/Keck. We demonstrate that scale-invariance constrains the two-field trajectory such that the effective dynamics are that of a single field, resulting in vanishing entropy perturbations and protecting the model from destabilization effects. We derive tight upper limits on the non-minimal coupling strength, excluding conformal coupling at high significance. By explicitly sampling over them, we demonstrate an overall insensitivity to initial conditions. We argue that the model predicts a minimal level of primordial tensor modes set by r ≳ 0.003, well within the reach of next-generation CMB experiments. These will therefore provide a litmus test of scale-invariant inflation, and we comment on the possibility of distinguishing the model from Starobinsky and α-attractor inflation. Overall, we argue that scale-invariant inflation is in excellent health, and possesses features which make it an interesting benchmark for tests of inflation from future CMB data.

The cosmological collider in R 2 inflation

Starobinsky's R 2 inflation manifests a best-fit scenario for the power spectrum of primordial density fluctuations. Observables derived from the slow-roll picture of the R 2 model in the Einstein frame relies on the conformal transformation of the metric, which inevitably induces a unique exponential-type couplings of the rolling scalaron with all matter fields during inflation. The “large-field” nature of the R 2 model further invokes non-negligible time and scale dependence to the matter sector through such an exponential coupling, modifying not only the dynamics of matter perturbations on superhorizon scales but also their decay rates. In this work, we identify the simplest observable of the cosmological collider physics built in the background of R 2 inflation, focusing on the so-called “quantum primordial clock” signals created by the non-local propagation of massive scalar perturbations. Our numerical formalism based on the unique conformal coupling can have extended applications to (quasi-)single-field inflationary models with non-trivial couplings to gravity or models that originated from the f(R) modification of gravity.

The Na+-translocating NADH:quinone oxidoreductase (Na+-NQR): Physiological role, structure and function of a redox-driven, molecular machine

Many bacterial processes are powered by the sodium motive force (smf) and in case of pathogens, the smf contributes to virulence. Vibrio cholerae, the causative agent of Cholera disease, possesses a Na+-translocating NADH:quinone oxidoreductase (NQR), a six-subunit membrane protein assembly. The 3D structure of NQR revealed the arrangement of the six subunits NqrABCDEF, the position of all redox cofactors (four flavins, two [2Fe-2S] centers) and the binding sites for the substrates NADH (in NqrF) and ubiquinone (in NqrB). Upon oxidation of NADH, electrons are shuttled twice across the membrane, starting with cytoplasmic FADNqrF and electron transfer to the [2Fe2S] clusterNqrF and from there to an intra-membranous [2Fe-2S] clusterNqrDE, periplasmic FMNNqrC, FMNNqrB and from there to riboflavinNqrB. This riboflavin is located at the cytoplasmic entry site of the sodium channel in NqrB, and it donates electrons to ubiquinone-8 positioned at the cytoplasmic side of NqrB. Targeting the substrate binding sites of NQR is a promising strategy to identify new inhibitors against many bacterial pathogens. Detailed structural information on the binding mode of natural inhibitors and small molecules in the active sites of NQR is now available, paving the way for the development of new antibiotics. The NQR shows different conformations as revealed in recent cryo-EM and crystallographic studies combined with spectroscopic analyses. These conformations represent distinct steps in the catalytic cycle. Considering the structural and functional data available, we propose a mechanism of Na+-NQR based on conformational coupling of electron transfer and Na+ translocation reaction steps.

Role of cosmological gravitational particle production in baryogenesis

We investigate the generation of the baryon asymmetry within the framework of cosmological gravitational particle production, employing the Bogoliubov approach. We examine two well-known baryogenesis scenarios, namely baryogenesis in grand unified theories (GUT) and leptogenesis, while considering reheating temperatures sufficiently low for thermal processes to be negligible. Considering α–attractor T-models for the inflaton potential, we demonstrate that GUT baryogenesis from scalar decays can be successful across a large range of conformal couplings with gravity, without necessitating substantial levels of CP violation. In the case of leptogenesis, we find that the reheating temperature should be TRH≲106 GeV for right-handed neutrino masses M1≲6×1012 GeV to generate the observed asymmetry. Published by the American Physical Society 2024

Conformal growth of B/N modified graphene on metal strings by chemical vapor deposition for robust protection

For string-plucked instruments, the metal strings are always subjected to sweating corrosion and ambient oxidation during the daily use. A specifically thin protection coating, which will not alter its timbre and tone, is highly desirable. In this study, a two-dimensional (2D) coating of B/N-modified graphene (BNG) films is proposed to deposit on the surface of Cu alloy strings via chemical vapor deposition (CVD) as a corrosion barrier. Through providing appropriate amount of B/N dopant, conformal growth of BNG films on Cu alloy string substrate with sharp step-terraces topography can be optimized, and largely improving its robust anti-corrosion performance in both short-term electrochemical and long-term ambient corrosion tests for evaluation. On one hand, the conformal coupling between in-situ grown BNG and Cu alloy string can form a strong interaction which limits the interfacial diffusion of corrosive species. On the other hand, once the defects oxidation is initialized, B/N modified graphene can reduce its conductivity, then suppressing the electrochemical corrosion in the long-term protection. With the insights and understanding of in-situ coating method and the enhanced anti-corrosion mechanisms, this work will extend the potential applications of 2D materials as an atomic-thick protectioncoating for some special devices and instruments.

Conformal and Non-Minimal Couplings in Fractional Cosmology

Fractional differential calculus is a mathematical tool that has found applications in the study of social and physical behaviors considered “anomalous”. It is often used when traditional integer derivatives models fail to represent cases where the power law is observed accurately. Fractional calculus must reflect non-local, frequency- and history-dependent properties of power-law phenomena. This tool has various important applications, such as fractional mass conservation, electrochemical analysis, groundwater flow problems, and fractional spatiotemporal diffusion equations. It can also be used in cosmology to explain late-time cosmic acceleration without the need for dark energy. We review some models using fractional differential equations. We look at the Einstein–Hilbert action, which is based on a fractional derivative action, and add a scalar field, ϕ, to create a non-minimal interaction theory with the coupling, ξRϕ2, between gravity and the scalar field, where ξ is the interaction constant. By employing various mathematical approaches, we can offer precise schemes to find analytical and numerical approximations of the solutions. Moreover, we comprehensively study the modified cosmological equations and analyze the solution space using the theory of dynamical systems and asymptotic expansion methods. This enables us to provide a qualitative description of cosmologies with a scalar field based on fractional calculus formalism.

Graph-based algorithms to dissect long-distance water-mediated H-bond networks for conformational couplings in GPCRs.

Changes in structure and dynamics elicited by agonist ligand binding at the extracellular side of G protein coupled receptors (GPCRs) must be relayed to the cytoplasmic G protein binding side of the receptors. To decipher the role of water-mediated hydrogen-bond networks in this relay mechanism, we have developed graph-based algorithms and analysis methodologies applicable to datasets of static structures of distinct GPCRs. For a reference dataset of static structures of bovine rhodopsin solved at the same resolution, we show that graph analyses capture the internal protein-water hydrogen-bond network. The extended analyses of static structures of rhodopsins and opioid receptors suggest a relay mechanism whereby inactive receptors have in place much of the internal core hydrogen-bond network required for long-distance relay of structural change, with extensive local H-bond clusters observed in structures solved at high resolution and with internal water molecules.

Weyl–invariant scalar–tensor gravities from purely metric theories

We describe a method to generate scalar–tensor theories with Weyl symmetry, starting from arbitrary purely metric higher derivative gravity theories. The method consists in the definition of a conformally-invariant metric g^μν\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\hat{g}_{\\mu \ u }$$\\end{document}, that is a rank (0,2)-tensor constructed out of the metric tensor and the scalar field. This new object has zero conformal weight and is given by ϕ2/Δgμν\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\phi ^{2/\\Delta }g_{\\mu \ u }$$\\end{document}, where (-Δ)\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$(-\\Delta )$$\\end{document} is the conformal dimension of the scalar. As gμν\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$g_{\\mu \ u }$$\\end{document} has conformal dimension of 2, the resulting tensor is trivially a conformal invariant. Then, the generated scalar–tensor theory, which we call the Weyl uplift of the original purely metric theory, is obtained by replacing the metric by g^μν\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\\hat{g}_{\\mu \ u }$$\\end{document} in the action that defines the original theory. This prescription allowed us to define the Weyl uplift of theories with terms of higher order in the Riemannian curvature. Furthermore, the prescription for scalar–tensor theories coming from terms that have explicit covariant derivatives in the Lagrangian is discussed. The same mechanism can also be used for the derivation of the equations of motion of the scalar–tensor theory from the original field equations in the Einstein frame. Applying this method of Weyl uplift allowed us to reproduce the known result for the conformal scalar coupling to Lovelock gravity and to derive that of Einsteinian cubic gravity. Finally, we show that the cancellation of the volume divergences in the theory given by the conformal scalar coupling to Einstein–Anti-de Sitter gravity is achieved by the Weyl uplift of the original theory augmented by counterterms, which is relevant in the framework of conformalrenormalization.

Signal integration in chemoreceptor complexes

Motile bacteria use large receptor arrays to detect chemical and physical stimuli in their environment, process this complex information, and accordingly bias their swimming in a direction they deem favorable. The chemoreceptor molecules form tripod-like trimers of receptor dimers through direct contacts between their cytoplasmic tips. A pair of trimers, together with a dedicated kinase enzyme, form a core signaling complex. Hundreds of core complexes network to form extended arrays. While considerable progress has been made in revealing the hierarchical structure of the array, the molecular properties underlying signal processing in these structures remain largely unclear. Here we analyzed the signaling properties of nonnetworked core complexes in live cells by following both conformational and kinase control responses to attractant stimuli and to output-biasing lesions at various locations in the receptor molecule. Contrary to the prevailing view that individual receptors are binary two-state devices, we demonstrate that conformational coupling between the ligand binding and the kinase-control receptor domains is, in fact, only moderate. In addition, we demonstrate communication between neighboring receptors through their trimer-contact domains that biases them to adopt similar signaling states. Taken together, these data suggest a view of signaling in receptor trimers that allows significant signal integration to occur within individual core complexes.

Dynamic and relativistic effects on Pt-Pt indirect spin-spin coupling in aqueous solution studied by abinitio molecular dynamics and two- vs four-component density functional NMR calculations.

Treating 195Pt nuclear magnetic resonance parameters in solution remains a considerable challenge from a quantum chemistry point of view, requiring a high level of theory that simultaneously takes into account the relativistic effects, the dynamic treatment of the solvent-solute system, and the dynamic electron correlation. A combination of Car-Parrinello molecular dynamics (CPMD) and relativistic calculations based on two-component zeroth order regular approximation spin-orbit Kohn-Sham (2c-ZKS) and four-component Dirac-Kohn-Sham (4c-DKS) Hamiltonians is performed to address the solvent effect (water) on the conformational changes and JPtPt1 coupling. A series of bridged PtIII dinuclear complexes [L1-Pt2(NH3)4(Am)2-L2]n+ (Am = α-pyrrolidonate and pivalamidate; L = H2O, Cl-, and Br-) are studied. The computed Pt-Pt coupling is strongly dependent on the conformational dynamics of the complexes, which, in turn, is correlated with the trans influence among axial ligands and with the angle N-C-O from the bridging ligands. The J-coupling is decomposed in terms of dynamic contributions. The decomposition reveals that the vibrational and explicit solvation contributions reduce JPtPt1 of diaquo complexes (L1 = L2 = H2O) in comparison to the static gas-phase magnitude, whereas the implicit solvation and bulk contributions correspond to an increase in JPtPt1 in dihalo (L1 = L2 = X-) and aquahalo (L1 = H2O; L2 = X-) complexes. Relativistic treatment combined with CPMD shows that the 2c-ZKS Hamiltonian performs as well as 4c-DKS for the JPtPt1 coupling.

Comprehensive Analysis of Coupled Proline Cis-Trans States in Bradykinin Using ωBP-REMD Simulations.

It is well-known that proline (Pro) cis-trans isomerization plays a decisive role in the folding and stabilization of proteins. The conformational coupling between isomerization states of different Pro residues in proteins during conformational adaptation processes is not well understood. In the present work, we investigate the coupled cis-trans isomerization of three Pro residues using bradykinin (BK), a partially unstructured nonapeptide hormone, as a model system. We use a recently developed enhanced-sampling molecular dynamics method (ω-bias potential replica exchange molecular dynamics; ωBP-REMD) that allows us to exhaustively sample all combinations of Pro isomer states and obtain converged probability densities of all eight state combinations within 885 ns ωBP-REMD simulations. In agreement with experiment, the all-trans state is seen to be the preferred isomer of zwitterionic aqueous BK. In about a third of its structures, this state presents the characteristic C-terminal β-turn conformation; however, other isomer combinations also contribute significantly to the structural ensemble. Unbiased probabilities can be projected onto the peptide bond dihedral angles of the three Pro residues. This unveils the interdependence of the individual Pro isomerization states, i.e., a possible coupling of the different Pro isomers. The cis/trans equilibrium of a Pro residue can change by up to 2.5 kcal·mol-1, depending on the isomerization state of other Pro residues. For example, for Pro7, the simulations indicate that its cis state becomes favored compared to its trans state when Pro2 is switched from the trans state to the cis state. Our findings demonstrate the efficiency of the ωBP-REMD methodology and suggest that the coupling of Pro isomerization states may play an even more decisive role in larger folded proteins subject to more conformational restraints.

Structural basis of DNA recognition of the Campylobacter jejuni CosR regulator.

Campylobacter jejuni has emerged as an antibiotic-resistant threat worldwide. CosR is an essential regulator for this bacterium and is important for Campylobacter adaptation to various stresses. Here, we describe the structural basis of CosR binding to target DNA as determined by cryo-electron microscopy and X-ray crystallography. Since CosR is a potential target for intervention, our studies may facilitate the development of novel therapeutics to combat C. jejuni infection.

Exploring conformational landscapes and binding mechanisms of convergent evolution for the SARS-CoV-2 spike Omicron variant complexes with the ACE2 receptor using AlphaFold2-based structural ensembles and molecular dynamics simulations.

In this study, we combined AlphaFold-based approaches for atomistic modeling of multiple protein states and microsecond molecular simulations to accurately characterize conformational ensembles evolution and binding mechanisms of convergent evolution for the SARS-CoV-2 spike Omicron variants BA.1, BA.2, BA.2.75, BA.3, BA.4/BA.5 and BQ.1.1. We employed and validated several different adaptations of the AlphaFold methodology for modeling of conformational ensembles including the introduced randomized full sequence scanning for manipulation of sequence variations to systematically explore conformational dynamics of Omicron spike protein complexes with the ACE2 receptor. Microsecond atomistic molecular dynamics (MD) simulations provide a detailed characterization of the conformational landscapes and thermodynamic stability of the Omicron variant complexes. By integrating the predictions of conformational ensembles from different AlphaFold adaptations and applying statistical confidence metrics we can expand characterization of the conformational ensembles and identify functional protein conformations that determine the equilibrium dynamics for the Omicron spike complexes with the ACE2. Conformational ensembles of the Omicron RBD-ACE2 complexes obtained using AlphaFold-based approaches for modeling protein states and MD simulations are employed for accurate comparative prediction of the binding energetics revealing an excellent agreement with the experimental data. In particular, the results demonstrated that AlphaFold-generated extended conformational ensembles can produce accurate binding energies for the Omicron RBD-ACE2 complexes. The results of this study suggested complementarities and potential synergies between AlphaFold predictions of protein conformational ensembles and MD simulations showing that integrating information from both methods can potentially yield a more adequate characterization of the conformational landscapes for the Omicron RBD-ACE2 complexes. This study provides insights in the interplay between conformational dynamics and binding, showing that evolution of Omicron variants through acquisition of convergent mutational sites may leverage conformational adaptability and dynamic couplings between key binding energy hotspots to optimize ACE2 binding affinity and enable immune evasion.

Graph-based analysis of H-bond networks and unsupervised learning reveal conformational coupling in prion peptide segments.

In this study, we employed a comprehensive computational approach to investigate the physical chemistry of the water networks surrounding hydrated peptide segments, as derived from molecular dynamics simulations. Our analysis uncovers a complex interplay of direct and water-mediated hydrogen bonds that intricately weave through the peptides. We demonstrate that these hydrogen bond networks encode critical information about the peptides' conformational behavior, with the dimensionality of these networks showing sensitivity to the peptides' conformations. Additionally, we estimated the free-energy landscape of the peptides across various conformations, revealing that their structures are predominantly characterized by unfolded, partially folded, and folded configurations, resulting in broad and rugged free-energy surfaces due to the numerous degrees of freedom contributed by the surrounding solvent. Importantly, the structured nature of this free-energy landscape becomes obscured when conventional collective variables, such as the number of hydrogen bonds, are used. Our findings provide new insights into the molecular mechanisms that couple protein and solvent degrees of freedom, highlighting their significance in the functioning of biological systems.

Higgs-induced screening mechanisms in scalar-tensor theories.

We consider the theory of a light conformally coupled scalar field, that is, one that is coupled directly to the Ricci scalar of the gravitational sector. This theory can be written equivalently as one of a light scalar that is coupled to the Standard Model of particle physics with a particular combination of Higgs-portal couplings. When the conformal coupling function contains terms that are linear and quadratic in the conformally coupled scalar, we find that the effective mass of the light propagating mode and its coupling to matter fields, obtained after expanding around a minimum of the classical potential, depend on the energy density of the background environment. This is despite the absence of nonlinear terms in the original equationof motion for the light conformally coupled field. Instead, we find that the nonlinearities of the prototype Higgs potential are communicated to the light mode. In this way, we present a novel realization of screening mechanisms, in which light degrees of freedom coupled to the Standard Model are able to avoid experimental constraints through environmental and thin-shelleffects.

Non-Noetherian conformal scalar fields

Recently, an extension of the standard four-dimensional scalar conformal action, yielding a second-order field equation that remains conformally invariant, was proposed. In spite of this, the corresponding action is not invariant under conformal transformations and this motivates us to define the notion of non-Noetherian conformal scalar field. In this article, we go further by determining the most general action in four dimensions that gives rise to a non-Noetherian conformal scalar field satisfying a second-order equation. This task is achieved by using the solution to the inverse problem of the calculus of variations. Surprisingly enough, the standard equation is shown to be extended by a non-Noetherian conformal piece involving a nonminimal coupling with a very particular combination of squared curvature terms, which is none other than the one defining the so-called Critical Gravity. We also prove that the most general second-order Euler–Lagrange equation for a conformal scalar field involves additional Noetherian conformal nonminimal couplings defined by an arbitrary function of the Weyl tensor. The recently proposed non-Noetherian conformal extension is recovered as a particular example of this function.

Orbital precession and hidden symmetries in scalar-tensor theories

We revisit the connection between relativistic orbital precession, the Laplace-Runge-Lenz symmetry, and the t-channel discontinuity of scattering amplitudes. Applying this to scalar-tensor theories of gravity, we compute the conservative potential and orbital precession induced by both conformal/disformal-type couplings at second Post-Minkowskian order (𝒪(GN 2)), complementing the known third/first order Post-Newtonian results. There is a particular tuning of the conformal coupling for which the precession vanishes at leading PN order, and we show that this coincides with the emergence of a Laplace-Runge-Lenz symmetry and a corresponding soft behaviour of the amplitude. While a single scalar field inevitably breaks this symmetry at higher PN orders, certain supersymmetric extensions have recently been shown to have an exact Laplace-Runge-Lenz symmetry and therefore classical orbits do not precess at any PN order. This symmetry can be used to relate scattering amplitudes at different loop orders, and we show how this may be used to bootstrap the (classically relevant part of the) three-loop 2 → 2 scattering of charged black holes in 𝒩 = 8 supergravity from existing two-loop calculations.

Higgs inflation at the pole

We propose a novel possibility for Higgs inflation where the perturbative unitarity below the Planck scale is ensured by construction and the successful predictions for inflation are accommodated. The conformal gravity coupling for the Higgs field leads to the proximity of the effective Planck mass to zero in the Jordan frame during inflation, corresponding to a pole in the Higgs kinetic term in the Einstein frame. Requiring the Higgs potential to vanish at the conformal pole in the effective theory in the Jordan frame, we make a robust prediction of the successful Higgs inflation. For a successful Higgs inflation at the pole, we take the running quartic coupling for the Higgs field to be small enough at the inflation scale, being consistent with the low-energy data, but we need a nontrivial extension of the SM with extra scalar or gauge fields in order to keep the running Higgs quartic coupling small during inflation. Performing the perturbative analysis of reheating with the known couplings of the SM particles to the Higgs boson, we show that a concrete realization of the Higgs pole inflation can be pinned down by the reheating processes with a general equation of state for the Higgs inflaton. We illustrate some extensions of the simple Higgs pole inflation to the general pole expansions, the running Higgs quartic coupling in the Standard Model and its extension with a singlet scalar field, a supergravity embedding of the Higgs pole inflation.

Engineering of Tunable Allosteric-like Fluorogenic Protein Sensors.

Optical protein sensors that enable detection of relevant biomolecules of interest play central roles in biological research. Coupling fluorescent reporters with protein sensing units has enabled the development of a wide range of biosensors that recognize analytes with high selectivity. In these sensors, analyte recognition induces a conformational change in the protein sensing unit that can modulate the optical signal of the fluorescent reporter. Here, we explore various designs for the creation of tunable allosteric-like fluorogenic protein sensors through incorporation of sensing protein units within the chemogenetic fluorescence-activating and absorption-shifting tag (FAST) that selectively binds and stabilizes the fluorescent state of 4-hydroxybenzylidene rhodanine (HBR) analogs. Conformational coupling allowed us to design analyte-responsive optical protein sensors through allosteric-like modulation of fluorogen binding.