ArticleBiophysical journal2009
Fluorescence fluctuation spectroscopy of mCherry in living cells.
Article in Biophysical journal, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 47 papers.
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Who cites it
47 citing papers in PubMed.
- Ultrafast Dynamics of rsCherryRev in Comparison with mCherry Reflecting Its Reversive Switchability.The journal of physical chemistry. B · 2026Article
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- Particle-based phasor-FLIM-FRET resolves protein-protein interactions inside single viral particles.Biophysical reports · 2023Article
- Benchmarking of novel green fluorescent proteins for the quantification of protein oligomerization in living cells.PloS one · 2023Article
- Quantification of Dark Protein Populations in Fluorescent Proteins by Two-Color Coincidence Detection and Nanophotonic Manipulation.The journal of physical chemistry. B · 2022Article
- Analysis and validation of silica-immobilised BST polymerase in loop-mediated isothermal amplification (LAMP) for malaria diagnosis.Analytical and bioanalytical chemistry · 2022Article
- Partitioning of ribonucleoprotein complexes from the cellular actin cortex.Science advances · 2022Article
- Intracellular imaging of metmyoglobin and oxygen using new dual purpose probe EYFP-Myoglobin-mCherry.Journal of biophotonics · 2022Article
- Selective Magnetic Nanoheating: Combining Iron Oxide Nanoparticles for Multi-Hot-Spot Induction and Sequential Regulation.Nano letters · 2021Article
- Number and Brightness Analysis: Visualization of Protein Oligomeric State in Living Cells.Advances in experimental medicine and biology · 2021Article
- Developing Analysis Protocols for Monitoring Intracellular Oxygenation Using Fluorescence Lifetime Imaging of Myoglobin-mCherry.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Real-time detection of somatic hybrid cells during electrofusion of carrot protoplasts with stably labelled mitochondria.Scientific reports · 2020Article
- Direct imaging of liquid domains in membranes by cryo-electron tomography.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Dengue Virus Capsid Protein Dynamics Reveals Spatially Heterogeneous Motion in Live-Infected-Cells.Scientific reports · 2020Article
- Differentiating Luminal and Membrane-Associated Nuclear Envelope Proteins.Biophysical journal · 2020Article
- Sensitive Detection of Protein Binding to the Plasma Membrane with Dual-Color Z-Scan Fluorescence.Biophysical journal · 2020Article
- Identifying Heteroprotein Complexes in the Nuclear Envelope.Biophysical journal · 2020Article
- Two-Color Spatial Cumulant Analysis Detects Heteromeric Interactions between Membrane Proteins.Biophysical journal · 2019Article
Corrections and comments
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Authors and funding
3 authors.
Funding
Abstract
The red fluorescent protein mCherry is of considerable interest for fluorescence fluctuation spectroscopy (FFS), because the wide separation in color between mCherry and green fluorescent protein provides excellent conditions for identifying protein interactions inside cells. This two-photon study reveals that mCherry exists in more than a single brightness state. Unbiased analysis of the data needs to account for the presence of multiple states. We introduce a two-state model that successfully describes the brightness and fluctuation amplitude of mCherry. The properties of the two states are characterized by FFS and fluorescence lifetime experiments. No interconversion between the two states was observed over the experimentally probed timescales. The effect of fluorescence resonance energy transfer between enhanced green fluorescent protein (EGFP) and mCherry is incorporated into the two-state model to describe protein hetero-oligomerization. The model is verified by comparing the predicted and measured brightness and fluctuation amplitude of several fusion proteins that contain mCherry and EGFP. In addition, hetero-fluorescence resonance energy transfer between mCherry molecules in different states is detected, but its influence on FFS parameters is small enough to be negligible. Finally, the two-state model is applied to study protein oligomerization in living cells. We demonstrate that the model successfully describes the homodimerization of nuclear receptors. In addition, we resolved a mixture of interacting and noninteracting proteins labeled with EGFP and mCherry. These results provide the foundation for quantitative applications of mCherry in FFS studies.
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