Bioluminescent proteins are used in a plethora of analytical methods, from ultrasensitive assay development to the in vivo imaging of cellular processes. This article reviews the most pertinent current bioluminescent-protein-based technologies and suggests the future direction of this vein of research. (To listen to a podcast about this feature, please go to the Analytical Chemistry multimedia page at pubs.acs.org/page/ancham/audio/index.html .).
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http://dx.doi.org/10.1021/ac9007286 | DOI Listing |
Methods Mol Biol
January 2025
Institute of Neurophysiology and NeuroCure Cluster of Excellence, Charité Universitätsmedizin Berlin, Berlin, Germany.
The bimolecular fluorescence complementation (BiFC) technique is a powerful tool for visualizing protein-protein interactions in vivo. It involves genetically fused nonfluorescent fragments of green fluorescent protein (GFP) or its variants to the target proteins of interest. When these proteins interact, the GFP fragments come together, resulting in the reconstitution of a functional fluorescent protein complex that can be observed using fluorescence microscopy.
View Article and Find Full Text PDFPLoS One
January 2025
Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
Exosomes are natural membrane-enclosed nanovesicles (30-150 nm) involved in cell-cell communication. Recently, they have garnered considerable interest as nanocarriers for the controlled transfer of therapeutic agents to cells. Here, exosomes were derived from bone marrow mesenchymal stem cells using three different isolation methods.
View Article and Find Full Text PDFAnal Chim Acta
February 2025
Joint Drug Development and Innovation Centre for Neurological Disorders of Lanzhou University-China National Biotec Group-Lanzhou Biotechnology Development Co., School of Pharmacy, Lanzhou University, Lanzhou, Gansu, 730000, PR China; MOE Frontiers Science Center for Rare Isotopes, Lanzhou University, Lanzhou, Gansu, 730000, PR China. Electronic address:
Background: Botulinum neurotoxin type A (BoNT/A) is the most potent and prevalent neurotoxin known to cause botulism, and is also widely used in medical and cosmetic applications. The detection of BoNT/A is of great significance for botulism diagnosis and drug potency determination. Currently, the mouse bioassay (MBA) has long been the gold standard method but has disadvantages of ethical concerns, long testing duration, and high costs.
View Article and Find Full Text PDFInt J Mol Sci
December 2024
Faculty of Pharmacy, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Osaka 920-1192, Japan.
G protein-coupled receptors (GPCRs) are essential cell surface proteins involved in transducing extracellular signals into intracellular responses, regulating various physiological processes. This study validated the use of the Tango assay, a sensitive method for detecting GPCR activation, in Schneider 2 (S2) cells, focusing on the human Dopamine Receptor D4 (DRD4). Plasmids encoding the LexA-tagged human DRD4 receptor and a luciferase reporter were co-transfected into S2 cells and stimulated with dopamine.
View Article and Find Full Text PDFInt J Mol Sci
December 2024
Shanghai Key Laboratory of Metabolic Remodeling and Health, Institute of Metabolism and Integrative Biology, Fudan University, Shanghai 200438, China.
Chaperone-mediated autophagy (CMA) is a selective autophagic pathway responsible for degrading cytoplasmic proteins within lysosomes. Monitoring CMA flux is essential for understanding its functions and molecular mechanisms but remains technically complex and challenging. In this study, we developed a pH-resistant probe, KFERQ-Gamillus, by screening various green fluorescent proteins.
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