ArticleBiophysical journal2012
Correlating calcium binding, Förster resonance energy transfer, and conformational change in the biosensor TN-XXL.
Article in Biophysical journal, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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Who cites it
25 citing papers in PubMed, 47 citations in OpenAlex.
- Calcium Buffering in Astrocytes and Its Relevance for Experimental Data Interpretation and Computational Modeling.Journal of neurochemistry · 2026Review
- FLIMB: fluorescence lifetime microendoscopy for metabolic and functional imaging of femoral marrow at subcellular resolution.Biomedical optics express · 2025Article
- Next-Generation Genetically Encoded Fluorescent Biosensors Illuminate Cell Signaling and Metabolism.Annual review of biophysics · 2024Review
- Calcium imaging: a technique to monitor calcium dynamics in biological systems.Physiology and molecular biology of plants : an international journal of functional plant biology · 2023Review
- Sub-millisecond conformational dynamics of the ACommunications biology · 2023Article
- Combined FRET-FLIM and NAD(P)H FLIM to Analyze B Cell Receptor Signaling Induced Metabolic Activity of Germinal Center B Cells In Vivo.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Structural Analysis of a Genetically Encoded FRET Biosensor by SAXS and MD Simulations.Sensors (Basel, Switzerland) · 2021Article
- Article
- β1-Integrin- and KV1.3 channel-dependent signaling stimulates glutamate release from Th17 cells.The Journal of clinical investigation · 2020Article
- Calcium-sensing receptor regulates intestinal dipeptide absorption via CaPhysiological reports · 2020Article
- Dynamic tuning of FRET in a green fluorescent protein biosensor.Science advances · 2019Article
- mCerulean3-Based Cameleon Sensor to Explore Mitochondrial CaiScience · 2019Article
- Integrated Strategies to Gain a Systems-Level View of Dynamic Signaling Networks.Methods in enzymology · 2017Article
- A Guide to Fluorescent Protein FRET Pairs.Sensors (Basel, Switzerland) · 2016Review
- Effect of Ca²⁺ on the steady-state and time-resolved emission properties of the genetically encoded fluorescent sensor CatchER.The journal of physical chemistry. B · 2015Article
- Article
- Biosensor architectures for high-fidelity reporting of cellular signaling.Biophysical journal · 2014Article
- Genetically encoded fluorescent biosensors for live-cell visualization of protein phosphorylation.Chemistry & biology · 2014Review
- Optimized ratiometric calcium sensors for functional in vivo imaging of neurons and T lymphocytes.Nature methods · 2014Article
- Putting a finishing touch on GECIs.Frontiers in molecular neuroscience · 2014Review
Corrections and comments
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Authors and funding
9 authors at 4 institutions in 1 country.
Funding
Abstract
Genetically encoded calcium indicators have become instrumental in imaging signaling in complex tissues and neuronal circuits in vivo. Despite their importance, structure-function relationships of these sensors often remain largely uncharacterized due to their artificial and multimodular composition. Here, we describe a combination of protein engineering and kinetic, spectroscopic, and biophysical analysis of the Förster resonance energy transfer (FRET)-based calcium biosensor TN-XXL. Using fluorescence spectroscopy of engineered tyrosines, we show that two of the four calcium binding EF-hands dominate the FRET output of TN-XXL and that local conformational changes of these hands match the kinetics of FRET change. Using small-angle x-ray scattering and NMR spectroscopy, we show that TN-XXL changes from a flexible elongated to a rigid globular shape upon binding calcium, thus resulting in FRET signal output. Furthermore, we compare calcium titrations using fluorescence lifetime spectroscopy with the ratiometric approach and investigate potential non-FRET effects that may affect the fluorophores. Thus, our data characterize the biophysics of TN-XXL in detail and may form a basis for further rational engineering of FRET-based biosensors.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.