2025/04/17 by Shosei Imai, Saaya Hario, Carl Suerte +8 · 1 voice
Biochemistry, Genetics and Molecular Biology · Chemistry · #Advanced biosensing and bioanalysis techniques #Advanced Biosensing Techniques and Applications #Advanced Proteomics Techniques and Applications
paper · pdf · doi:10.1101/2025.04.17.649293
Abstract Pyruvate is the end-product of glycolysis and a central metabolite involved in many biochemical pathways. However, a lack of high-performance (i.e., Δ F / F 0 > 10) single fluorescent protein (FP)-based biosensors has hindered efforts to investigate the physiological role of pyruvate. Here, we present the GreenPy1 and ApplePy1 series, which are green FP (GFP)-based and red FP (RFP)-based pyruvate biosensors, respectively. Both series exhibit large fluorescence intensity change (Δ F / F 0 ∼ 20 to >40) and a range of affinities (10s of μM to several mM). We demonstrate the utility of these pyruvate biosensors for multicolor imaging of metabolite concentration changes in mammalian cells.