ArticleInternational journal of molecular sciences2021
Intracellular Ionic Strength Sensing Using NanoLuc.
Article in International journal of molecular sciences, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- E50A Mutation Increases the Bioluminescence Activity of picALuc.Biosensors · 2026Article
- Continuous Hypermutation and Evolution of Luciferase Variants.ACS chemical biology · 2026Article
- Differential Impact of Substrate Peptides on Interdomain Interactions in Severe Acute Respiratory Syndrome Coronavirus 2 Main Protease.Computational and structural biotechnology journal · 2026Article
- Article
- Bioluminescent enzymatic biosensors: ways to manage their characteristics.Biophysical reviews · 2025Review
- Comparative Analysis of Biochemical and Cellular Assay Conditions and the Need for a Buffer That Mimics Cytoplasmic Environments.Molecules (Basel, Switzerland) · 2025Review
- Coevolving residues distant from the ligand binding site are involved in GAF domain function.Communications chemistry · 2025Article
- Luminescence Fingerprint of Intracellular NIR-II Gold Nanocluster Transformation: Implications for Sensing and Imaging.ACS nano · 2025Article
- Physicochemical Perspective of Biological Heterogeneity.ACS physical chemistry Au · 2024Review
- A potential allosteric inhibitor of SARS-CoV-2 main protease (MFrontiers in molecular biosciences · 2024Article
- Interference of metal ions on the bioluminescent signal of firefly, Renilla, and NanoLuc luciferases in high-throughput screening assays.Frontiers in chemistry · 2024Article
- BRET-based biosensors for SARS-CoV-2 oligonucleotide detection.Frontiers in bioengineering and biotechnology · 2024Article
- TRIM21 and Fc-engineered antibodies: decoding its complex antibody binding mode with implications for viral neutralization.Frontiers in immunology · 2024Article
- Illuminating the mechanism and allosteric behavior of NanoLuc luciferase.Nature communications · 2023Article
- Functional Characterization of NovelInternational journal of molecular sciences · 2023Article
- S-Series Coelenterazine-Driven Combinatorial Bioluminescence Imaging Systems for Mammalian Cells.International journal of molecular sciences · 2023Article
- SARS-CoV-2 accessory proteins ORF7a and ORF3a use distinct mechanisms to down-regulate MHC-I surface expression.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Protegrin-2, a potential inhibitor for targeting SARS-CoV-2 main protease MComputational and structural biotechnology journal · 2023Article
- Reversal of the unique Q493R mutation increases the affinity of Omicron S1-RBD for ACE2.Computational and structural biotechnology journal · 2023Article
- Macromolecular assembly of bioluminescent protein nanoparticles for enhanced imaging.Materials today. Bio · 2022Article
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Authors and funding
4 authors.
Funding
Abstract
Intracellular ionic strength regulates myriad cellular processes that are fundamental to cellular survival and proliferation, including protein activity, aggregation, phase separation, and cell volume. It could be altered by changes in the activity of cellular signaling pathways, such as those that impact the activity of membrane-localized ion channels or by alterations in the microenvironmental osmolarity. Therefore, there is a demand for the development of sensitive tools for real-time monitoring of intracellular ionic strength. Here, we developed a bioluminescence-based intracellular ionic strength sensing strategy using the Nano Luciferase (NanoLuc) protein that has gained tremendous utility due to its high, long-lived bioluminescence output and thermal stability. Biochemical experiments using a recombinantly purified protein showed that NanoLuc bioluminescence is dependent on the ionic strength of the reaction buffer for a wide range of ionic strength conditions. Importantly, the decrease in the NanoLuc activity observed at higher ionic strengths could be reversed by decreasing the ionic strength of the reaction, thus making it suitable for sensing intracellular ionic strength alterations. Finally, we used an mNeonGreen-NanoLuc fusion protein to successfully monitor ionic strength alterations in a ratiometric manner through independent fluorescence and bioluminescence measurements in cell lysates and live cells. We envisage that the biosensing strategy developed here for detecting alterations in intracellular ionic strength will be applicable in a wide range of experiments, including high throughput cellular signaling, ion channel functional genomics, and drug discovery.
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Registered trials
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