Abstract: The present disclosure provides methods for identifying compounds that cause structural changes in a protein bound to an actin filament. The methods include the use of cells that include two actin-binding proteins, each labeled with a chromophore, and exposing the cells to a test compound. The method further includes detecting a change in fluorescence resonance energy transfer (FRET) between the chromophores.

Abstract: A protein kinase that includes a donor molecule and an acceptor molecule, methods of making the protein kinase, and methods of using the protein kinase are described. Measurement of the conformation of the kinase can be obtained using intramolecular FRET. The protein kinase can be used to, for example, identify conformational changes involved in kinase regulation, that is, as an allostery sensor; to identify kinase-binding molecules including, for example, kinase inhibitors; to identify cancer therapeutics; or for high-throughput screening.

Abstract: Provided herein are methods for identifying a compound that alters fluorescence resonance energy transfer (FRET) of a protein. In one embodiment, the method includes providing a target protein, where the target protein includes two heterologous domains, each domain having chromophores that together act as a FRET pair. In another embodiment, the method includes providing a target protein and a second protein, wherein the target protein includes a first heterologous domain including a chromophore, and the second protein includes a second heterologous domain including a chromophore, where the chromophores together act as a FRET pair. The method further includes contacting a sample including the target protein and optional second protein with a test compound to form a mixture, and measuring a fluorescence emission spectrum of the mixture during exposure to a light source.

Abstract: The present disclosure provides methods for identifying compounds that cause structural changes in a protein bound to an actin filament. The methods include the use of cells that include two actin-binding proteins, each labeled with a chromophore, and exposing the cells to a test compound. The method further includes detecting a change in fluorescence resonance energy transfer (FRET) between the chromophores.

Abstract: Methods for identifying a compound that alters fluorescence resonance energy transfer (FRET) of a protein. The methods include use of a genetically engineered cell that includes a target protein. The target protein includes one or more heterologous domains. In one embodiment, a target protein includes two heterologous domains, and in another embodiment, the target protein includes a heterologous domain and the cell further includes a second protein that includes a heterologous domain. A heterologous domain may include a chromophore or an amino acid to which a fluorescent dye attaches. The fluorescence lifetime of one or more chromophore, one or more fluorescent dye, or the combination thereof, is measured after contacting the cell with a compound A difference between the fluorescence lifetime in the presence of the test compound and the fluorescence lifetime in the absence of the test compound indicates that the test compound alters the FRET of the target protein.

Abstract: A protein kinase that includes a donor molecule and an acceptor molecule, methods of making the protein kinase, and methods of using the protein kinase are described. Measurement of the conformation of the kinase can be obtained using intramolecular FRET. The protein kinase can be used to, for example, identify conformational changes involved in kinase regulation, that is, as an allostery sensor; to identify kinase-binding molecules including, for example, kinase inhibitors; to identify cancer therapeutics; or for high-throughput screening.

Abstract: Provided herein are methods for identifying a compound that alters fluorescence resonance energy transfer (FRET) of a protein. In one embodiment, the method includes providing a target protein, where the target protein includes two heterologous domains, each domain having chromophores that together act as a FRET pair. In another embodiment, the method includes providing a target protein and a second protein, wherein the target protein includes a first heterologous domain including a chromophore, and the second protein includes a second heterologous domain including a chromophore, where the chromophores together act as a FRET pair. The method further includes contacting a sample including the target protein and optional second protein with a test compound to form a mixture, and measuring a fluorescence emission spectrum of the mixture during exposure to a light source.

Abstract: Provided herein are methods for identifying a compound that modulates a Ryanodine receptor (Ryr). Fluorescence resonance energy transfer between an FKBP bound to an RyR and fluorescent derivatives of RyR binding partners (e.g., calmodulin) or domain-peptide biosensors is used to provide a readout dependent on the RyR functional state. The methods permit measurement of RyR present in a permeabilized cell or in a purified membrane.

Abstract: Provided herein are methods for identifying a compound that alters fluorescence resonance energy transfer (FRET) of a protein. In one embodiment, the method includes providing a target protein, where the target protein includes two heterologous domains, each domain having chromophores that together act as a FRET pair. In another embodiment, the method includes providing a target protein and a second protein, wherein the target protein includes a first heterologous domain including a chromophore, and the second protein includes a second heterologous domain including a chromophore, where the chromophores together act as a FRET pair. The method further includes contacting a sample including the target protein and optional second protein with a test compound to form a mixture, and measuring a fluorescence emission spectrum of the mixture during exposure to a light source.

Abstract: Methods for identifying a compound that alters fluorescence resonance energy transfer (FRET) of a protein. The methods include use of a genetically engineered cell that includes a target protein. The target protein includes one or more heterologous domains. In one embodiment, a target protein includes two heterologous domains, and in another embodiment, the target protein includes a heterologous domain and the cell further includes a second protein that includes a heterologous domain. A heterologous domain may include a chromophore or an amino acid to which a fluorescent dye attaches. The fluorescence lifetime of one or more chromophore, one or more fluorescent dye, or the combination thereof, is measured after contacting the cell with a compound A difference between the fluorescence lifetime in the presence of the test compound and the fluorescence lifetime in the absence of the test compound indicates that the test compound alters the FRET of the target protein.

Abstract: Provided herein are methods for identifying a compound that modulates a Ryanodine receptor (Ryr). Fluorescence resonance energy transfer between an FKBP bound to an RyR and fluorescent derivatives of RyR binding partners (e.g., calmodulin) or domain-peptide biosensors is used to provide a readout dependent on the RyR functional state. The methods permit measurement of RyR present in a permeabilized cell or in a purified membrane.

Abstract: Provided herein are methods for identifying molecules capable of modulating SERCA, the SERCA-PLB complex or the microenvironment of the complex. An exemplary assay provided herein is fluorescence resonance energy transfer (FRET). Also provided herein are FRET assays that are optimized for high-throughput screening (HTS) for identifying small molecules that modulate SERCA or the SERCA-PLB complex. Further provided are kits for carrying out said methods for identifying molecules.

Abstract: Provided herein are methods for identifying molecules capable of modulating SERCA, the SERCA-PLB complex or the microenvironment of the complex. An exemplary assay provided herein is fluorescence resonance energy transfer (FRET). Also provided herein are FRET assays that are optimized for high-throughput screening (HTS) for identifying small molecules that modulate SERCA or the SERCA-PLB complex. Further provided are kits for carrying out said methods for identifying molecules.

Abstract: An exercise bar is rotatably mounted relative to a central support rod, the central rod including extension members extending coaxially and laterally of the bar for mounting within opposed vertical door frame portions. The bar includes an exterior shell rotatably mounted relative to an interior shell, wherein a modification of the invention includes a central rod portion of a first diameter less than that of spaced end portions to accommodate individuals of various sized hands for mounting onto the bar.