A novel FbFP-based biosensor toolbox for sensitive in vivo determination of intracellular pH

Journal of Biotechnology
Christian RupprechtThomas Drepper

Abstract

The intracellular pH is an important modulator of various bio(techno)logical processes such as enzymatic conversion of metabolites or transport across the cell membrane. Changes of intracellular pH due to altered proton distribution can thus cause dysfunction of cellular processes. Consequently, accurate monitoring of intracellular pH allows elucidating the pH-dependency of (patho)physiological and biotechnological processes. In this context, genetically encoded biosensors represent a powerful tool to determine intracellular pH values non-invasively and with high spatiotemporal resolution. We have constructed a toolbox of novel genetically encoded FRET-based pH biosensors (named Fluorescence Biosensors for pH or FluBpH) that utilizes the FMN-binding fluorescent protein EcFbFP as donor domain. In contrast to many fluorescent proteins of the GFP family, EcFbFP exhibits a remarkable tolerance towards acidic pH (pKa∼3.2). To cover the broad range of physiologically relevant pH values, three EYFP variants exhibiting pKa values of 5.7, 6.1 and 7.5 were used as pH-sensing FRET acceptor domains. The resulting biosensors FluBpH 5.7, FluBpH 6.1 and FluBpH 7.5 were calibrated in vitro and in vivo to accurately evaluate their pH indicator ...Continue Reading

Citations

Jun 27, 2018·Microbial Biotechnology·Alexander BollingerKarl-Erich Jaeger
Dec 11, 2019·International Journal of Molecular Sciences·Tiphaine Péresse, Arnaud Gautier
Oct 4, 2018·Mikrochimica Acta·Lude Wang, Yang Chen
Aug 19, 2020·Current Opinion in Microbiology·Béatrice Roche, Dirk Bumann
Nov 27, 2020·Frontiers in Bioengineering and Biotechnology·Longwei LiuYingxiao Wang
Apr 17, 2021·The Journal of Biological Chemistry·Katrin RöllenUlrich Krauss
May 1, 2021·Frontiers in Microbiology·Anaïs Biquet-BisquertAshley L Nord
Apr 4, 2019·The Journal of Physical Chemistry. B·Joshua D SlocumRalph Jimenez
Mar 14, 2019·ACS Sensors·Sandra BurgstallerRoland Malli
Dec 4, 2019·Bioconjugate Chemistry·Harun F OzbakirArnab Mukherjee
Jul 10, 2018·Chemical Reviews·Aba LosiAndreas Möglich
Apr 4, 2019·Metabolic Engineering·Volker F Wendisch

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