title: Firefly Luciferase-based Fusion Proteins and their Applications in Bioanalysis.
abstract: Firefly luciferase is widely used in molecular biology and bioanalytical systems as a reporter molecule due to the high quantum yield of the bioluminescence, availability of stable mutant forms of the enzyme with prescribed spectral characteristics and abundance of bacterial expression systems suitable for production of recombinant proteins in limitless quantities. In this review, we described fusion proteins of luciferase with biotin-binding domain and streptavidin, with proteins A and G, antibodies, with DNA- and RNA-binding proteins, as well as fusion proteins designed for BRET systems. The firefly luciferase-based fusion proteins are represented as an effective tool for the development of different bioanalytical systems such as (1) systems in which luciferase is attached to the surface of the target and the bioluminescence signal is detected from the specific complexes formed; (2) BRET-based systems, in which the specific interaction induces changes in the bioluminescence spectrum; and (3) systems that use modified or split luciferases, in which the luciferase activity changes under the action of the analyte. All these systems have wide application in biochemical analysis of physiologically important compounds, for the detection of pathogenic bacteria and viruses, for evaluation of protein-protein interactions, assaying of metabolites involved in cell communication and cell signaling.
authors: Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
authors: Daria V
authors: DV
authors: Smirnova
authors: Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia.
authors: Natalia N
authors: NN
authors: Ugarova
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015374:Biosensing Techniques; D004268:DNA-Binding Proteins / Q000378:metabolism; D004735:Energy Transfer; D048888:Fireflies / Q000201:enzymology*; D007074:Immunoglobulin G / Q000378:metabolism; D049409:Luciferases, Firefly / Q000235:genetics / Q000378:metabolism*; D049449:Luminescence; D049408:Luminescent Agents / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D011485:Protein Binding; D000072417:Protein Domains; D016601:RNA-Binding Proteins / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013205:Staphylococcal Protein A / Q000378:metabolism
chemical_list: D000906:Antibodies; D004268:DNA-Binding Proteins; D007074:Immunoglobulin G; D049408:Luminescent Agents; D016601:RNA-Binding Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D049409:Luciferases, Firefly