Dual enzyme activated fluorescein based fluorescent probe

Abstract

A simple dual analyte fluorescein-based probe ( PF3-Glc) was synthesised containing β-glucosidase ( β-glc) and hydrogen peroxide (H ₂O ₂) trigger units. The presence of β-glc, resulted in fragmentation of the parent molecule releasing glucose and the slightly fluorescent mono-boronate fluorescein ( PF3). Subsequently, in the presence of glucose oxidase (GOx), the released glucose was catalytically converted to D-glucono- δ-lactone, which produced H ₂O ₂ as a by-product. The GOx-produced H ₂O _2, resulted in classic H ₂O ₂-mediated boronate oxidation and the release of the highly emissive fluorophore, fluorescein. This unique cascade reaction lead to an 80-fold increase in fluorescence intensity.

Graphical abstract

Keywords

chemosensors / dual-activation / GOx / fluorescence / β-glucosidase / molecular logic

References

[1] Medina R, Owen G I. Glucose transporters: Expression, regulation and cancer. Journal of Biological Research (Thessaloniki), 2002, 35(1): 9–26

[2] Iadecola C. Sugar and Alzheimer’s disease: A bittersweet truth. Nature Neuroscience, 2015, 18(4): 477–478

[3] An Y, Varma V R, Varma S, Casanova R, Dammer E, Pletnikova O, Chia C W, Egan J M, Ferrucci L, Troncoso J, Levey A I, Lah J, Seyfried N T, Legido-Quigley C, O’Brien R, Thambisetty M. Evidence for brain glucose dysregulation in Alzheimer’s disease. Alzheimer’s & Dementia, 2018, 14(3): 318–329

[4] Körschen H G, Yildiz Y, Raju D N, Schonauer S, Bönigk W, Jansen V, Kremmer E, Kaupp U B, Wachten D. The non-lysosomal β-glucosidase GBA2 is a non-integral membrane-associated protein at the endoplasmic reticulum (ER) and Golgi. Journal of Biological Chemistry, 2012, 288(5): 12–20

[5] Zhou X, Huang Z, Yang H, Jiang Y, Wei W, Li Q, Mo Q, Lui J. β-Glucosidase inhibtion sensitises breast cancer to chemotherapy. Journal of Biomedicine and Pharmacotherapy, 2017, 91: 504–509

[6] Wong C M, Wong K H, Chen X D. Glucose oxidase: Natural occurrence, function, properties and industrial applications. Applied Microbiology and Biotechnology, 2008, 78(6): 927–938

[7] Ramanavicius A, Ryskevic N, Kausaite-Minkstimiene A, Bubniene U, Baleviciute I, Oztekin Y, Ramanaviciene A. Fluorescence study of glucose oxidase self-encapsulated within polypyrrole. Sensors and Actuators. B, Chemical, 2012, 171: 753–759

[8] Chan J, Dodani S C, Chang C J. Reaction-based small-molecule fluorescent probes for chemoselective bioimaging. Nature Chemistry, 2012, 4(12): 973–984

[9] Wu D, Sedgwick A C, Gunnlaugsson T, Akkaya E U, Yoon J, James T D. Fluorescent chemosensors: The past, present and future. Chemical Society Reviews, 2017, 46(23): 7105–7123

[10] Reeves E P, Lu H, Jacobs H L, Messina C G M, Bolsover S, Gabella G, Potma E O, Warley A, Roes J, Segal A W. Killing activity of neutrophils is mediated through activation of proteases by K Flux. Nature, 2002, 416(6878): 291–297

[11] Winterbourn C C. Reconciling the chemistry and biology of reactive oxygen species. Nature Chemical Biology, 2008, 4(5): 278–286

[12] Wang N N, Miller C J, Wang P, Waite T D. Quantitative determination of trace hydrogen peroxide in the presence of sulfide using the Amplex Red/horseradish peroxidase assay. Analytica Chimica Acta, 2017, 963: 61–67

[13] Quinlan C L, Perevoschikova I V, Goncalves R L S, Hey-Mogensen M, Brand M D. The determination and analysis of site-specific rates of mitochondrial reactive oxygen species production. Methods in Enzymology, 2013, 523: 189–217

[14] Wannajuk K, Jamkatoke M, Tuntulani T, Tomapatanaget B. Highly specific-glucose fluorescence sensing based on boronic anthraquinone derivatives via the GOx enzymatic reaction. Tetrahedron, 2012, 68(43): 8899–8904

[15] Sedgwick A C, Han H H, Gardiner J E, Bull S D, He X P, James T D. The development of a novel AND logic based fluorescence probe for the detection of peroxynitrite and GSH. Chemical Science (Cambridge), 2018, 9(15): 3672–3676

[16] Dickinson B C, Huynh C, Chang C J. A palette of fluorescent probes with varying emission colours for imaging hydrgoen peroxide signalling in living cells. Journal of the American Chemical Society, 2010, 132(16): 5906–5915

[17] Dickinson B C, Chang C J. A targetable fluorescent probe for imaging hydrogen peroxide in the mitochondria of living cells. Journal of the American Chemical Society, 2008, 130(30): 9638–9639

[18] Hong K H, Kim D I, Kwon H, Kim H J. A fluoresceinylcarbonate-based fluorescent probe for the sensitive detection of biothiols in a HEPES buffer and its cellular expression. RSC Advances, 2014, 4(2): 978–982