Abstract: Provided herein are systems, methods, and compositions for bioluminescence-triggered photocatalytic activation of molecular entities in a proximity-dependent manner, which can be actuated within biological systems. In particular, provided herein are bioluminescent proteins or complexes, luminophore substrates thereof, photocatalysts, and activatable molecular entities incorporating light-responsive moieties that restrict their activity; systems thereof; and methods for catalytically activating the activatable molecular entities via bioluminescence-triggered catalysis.

Abstract: Provided herein are systems, methods, and compositions for bioluminescence-triggered catalysis of bioorthogonal labeling chemistries in a proximity dependent manner. In particular, provided herein are bioluminescent proteins or complexes, luminophore substrates thereof, photocatalysts or photosensitizers, activatable labels, and systems thereof, and methods of catalytically activating the activatable labels via bioluminescence-triggered catalysis.

Abstract: Provided herein modified dehalogenases have extended surface loop regions that provide a location for internal fusion insertions and modulate binding interaction and activation of environmentally-sensitive chemistries.

Abstract: Described are substituted imidazo[1,2-a]pyrazine compounds, which are coelenterazine analogues, kits comprising the analogues, and methods of using the compounds for the detection of luminescence in luciferase-based assays. Also described are methods from making the compounds, such as a method using aminopyrazine acetophosphonates as synthesis intermediates.

Abstract: Provided herein are circularly-permuted (cp) dehalogenase variants that are capable of covalently binding to a haloalkyl ligand. In particular cp dehalogenase variants and peptides and polypeptides comprising split versions thereof that structurally assemble to form an active dehalogenase complexes are provided.

Abstract: Provided herein are systems and methods for characterizing target/ligand engagement. In particular, luciferase-labeled polypeptide targets are used to detect or quantify target/ligand engagement (e.g., within a cell or cell lysate).

Abstract: Provided herein are compositions and methods for detection and analysis of intracellular binding of a bioactive agent to a cellular target. In particular, bioactive agents tethered to fluorophores, cellular targets fused to bioluminescent reporters, and methods of detecting and analyzing the interaction of bioactive agents with cellular targets therewith.

Abstract: Compounds that may inhibit Oplophorus-derived luciferases are disclosed as well as compositions and kits comprising the compounds and methods of using the compounds.

Abstract: Provided herein are compounds, compositions, systems, and methods for photoactivatable labeling, which can be actuated within biological systems. In particular, compounds disclosed herein include vinyl-extended-aryl azide moieties that undergo photoactivation to generate reactive intermediates, which can form covalent linkages with biomolecules. The photoactivation can be conducted by a variety of mechanisms including ultraviolet (UV) irradiation, visible light irradiation, or energy transfer (e.g., from a photocatalyst). The compounds also include functional moieties that provide useful functionalities, for example detection of biomolecules, such as fluorophores, capture elements (e.g., biotin), or reactive moieties (e.g., click handles) and bifunctional moieties comprising a bioactive compound and a detection/capture element.

Abstract: Provided herein are systems and methods for characterizing target/ligand engagement. In particular, luciferase-labeled polypeptide targets are used to detect or quantify target/ligand engagement (e.g., within a cell or cell lysate).

Abstract: Provided herein are compositions and methods for the assembly of a bioluminescent complex from two or more non-luminescent (e.g., substantially non-luminescent) peptide and/or polypeptide units. In particular, bioluminescent activity is conferred upon a non-luminescent polypeptide via structural complementation with another, complementary non-luminescent peptide.

Abstract: Described are processes for preparing substituted imidazo[1,2-a]pyrazine compounds of formula (I): by reacting a compound of formula (xi): with carbonyldiimidazole to form the compound of formula (I).

Abstract: Provided herein are compositions and methods for detection and analysis of intracellular binding of a bioactive agent to a cellular target. In particular, bioactive agents tethered to fluorophores, cellular targets fused to bioluminescent reporters, and methods of detecting and analyzing the interaction of bioactive agents with cellular targets therewith.

Abstract: Compounds that may inhibit Oplophorus-derived luciferases are disclosed as well as compositions and kits comprising the compounds and methods of using the compounds.

Abstract: Provided herein are compositions and methods for stabilizing coelenterazine and analogs or derivatives thereof, and for improving the solubility and reconstitution efficiency of coelenterazine and analogs and derivatives thereof.

Abstract: Provided herein are systems and methods for characterizing target/ligand engagement. In particular, luciferase-labeled polypeptide targets are used to detect or quantify target/ligand engagement (e.g., within a cell or cell lysate).

Abstract: Provided herein are compositions and methods for stabilizing coelenterazine and analogs or derivatives thereof, and for improving the solubility and reconstitution efficiency of coelenterazine and analogs and derivatives thereof.

Abstract: Provided herein are compositions and methods for the assembly of a bioluminescent complex from two or more non-luminescent (e.g., substantially non-luminescent) peptide and/or polypeptide units. In particular, bioluminescent activity is conferred upon a non-luminescent polypeptide via structural complementation with another, complementary non-luminescent peptide.

Abstract: Provided herein are substantially non-luminescent peptide/polypeptide tags that are inserted internally within a protein of interest or between N-terminal and C-terminal peptides/polypeptides. Interaction of the internally-inserted tag with a complement polypeptide/peptide that is also substantially non-luminescent results in the formation a bioluminescent reporter complex.

Abstract: Provided herein are substantially non-luminescent peptide/polypeptide tags that are inserted internally within a protein of interest or between N-terminal and C-terminal peptides/polypeptides. Interaction of the internally-inserted tag with a complement polypeptide/peptide that is also substantially non-luminescent results in the formation a bioluminescent reporter complex.