Abstract—
Recently, fluorescent proteins have become an indispensable tool for in vivo visualization of various processes occurring in living systems, from individual organelles to whole organisms. They allow observing the expression of proteins, their localization, and are also suitable for monitoring biochemical processes in cells. As a rule, fluorescent proteins have pH-dependent spectral properties, which make it possible to design genetically encoded pH sensors on their basis for solving various biological problems. In this work, we have obtained a set of extracellular pH sensors based on the SypHer3S protein, which has pH sensitivity in the pH range from 7.4 to 9.0. For this purpose, chimeric SypHer3S constructs with various transmembrane domains of membrane receptors were created, allowing the targeting of this protein to the outer side of the cytoplasmic membrane. Their subcellular localization was characterized and the most successful version of a chimeric pH sensor for measuring the extracellular pH value was selected.
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ACKNOWLEDGMENTS
The authors are grateful to V.V. Belousov for providing the SypHer3S cDNA.
Funding
The study was carried out with the financial support of the Russian Foundation for Basic Research: project nos. 19-34-90177, 20-04-00880, 19-04-01042, 20-04-00959, 17-00-00486, 17-00-00489, 19-04-00815, 19-34-51034, 18-04-01369, 18-29-09166, and 20-34-90019.
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The article does not contain a description of research carried out by any of the authors of this work, involving humans or using animals as objects.
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Abbreviations: CIRL2, calcium-independent receptor for alpha-latrotoxin 2 (latrophilin 2); c-Met, hepatocyte growth factor receptor; ErbB2, epidermal growth factor receptor; IR, insulin receptor; IRR, insulin receptor-related receptor; PDGFR, platelet-derived growth factor receptor; SypHer3S, yellow GFP-like protein; TrkA, tropomyosin receptor kinase A.
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Orsa, A.N., Goryashchenko, A.S., Serova, O.V. et al. Generation and Characteristics of Genetically Encoded Fluorescent Sensors of Extracellular pH. Russ J Bioorg Chem 47, 1060–1065 (2021). https://doi.org/10.1134/S1068162021040178
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DOI: https://doi.org/10.1134/S1068162021040178