ArticleBiophysical journal2005
Unraveling protein-protein interactions in living cells with fluorescence fluctuation brightness analysis.
Article in Biophysical journal, 2005. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 42 citations in OpenAlex.
- Article
- Agonist-controlled competition of RAR and VDR nuclear receptors for heterodimerization with RXR is manifested in their DNA binding.The Journal of biological chemistry · 2023Article
- Identifying Heteroprotein Complexes in the Nuclear Envelope.Biophysical journal · 2020Article
- Auto-FPFA: An Automated Microscope for Characterizing Genetically Encoded Biosensors.Scientific reports · 2018Article
- Biochemical, Biophysical and Cellular Techniques to Study the Guanine Nucleotide Exchange Factor, GIV/Girdin.Current protocols in chemical biology · 2016Article
- Dopamine Receptor Signaling in MIN6 β-Cells Revealed by Fluorescence Fluctuation Spectroscopy.Biophysical journal · 2016Article
- Characterization of Ternary Protein Systems In Vivo with Tricolor Heterospecies Partition Analysis.Biophysical journal · 2016Article
- Quantifying protein-protein interactions of peripheral membrane proteins by fluorescence brightness analysis.Biophysical journal · 2014Article
- Interrelationship between cytoplasmic retroviral Gag concentration and Gag-membrane association.Journal of molecular biology · 2014Article
- The statistics of protein expression ratios for cellular fluorescence studies.European biophysics journal : EBJ · 2012Article
- The retinoid X receptors and their ligands.Biochimica et biophysica acta · 2012Review
- Characterization of cytoplasmic Gag-gag interactions by dual-color z-scan fluorescence fluctuation spectroscopy.Biophysical journal · 2011Article
- Heterospecies partition analysis reveals binding curve and stoichiometry of protein interactions in living cells.Proceedings of the National Academy of Sciences of the United States of America · 2010Article
- Pirenzepine promotes the dimerization of muscarinic M1 receptors through a three-step binding process.The Journal of biological chemistry · 2009Article
- Fluorescence fluctuation spectroscopy of mCherry in living cells.Biophysical journal · 2009Article
- Fluorescence fluctuation spectroscopy on viral-like particles reveals variable gag stoichiometry.Biophysical journal · 2009Article
- Fluorescence fluctuation analysis of Arabidopsis thaliana somatic embryogenesis receptor-like kinase and brassinosteroid insensitive 1 receptor oligomerization.Biophysical journal · 2008Article
- Pharmacology under the microscope: the use of fluorescence correlation spectroscopy to determine the properties of ligand-receptor complexes.Trends in pharmacological sciences · 2007Review
- Oligomerization of the EGF receptor investigated by live cell fluorescence intensity distribution analysis.Biophysical journal · 2007Article
- Determining the stoichiometry of protein heterocomplexes in living cells with fluorescence fluctuation spectroscopy.Proceedings of the National Academy of Sciences of the United States of America · 2007Article
Corrections and comments
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Fluorescence correlation spectroscopy is a potentially powerful tool for measuring protein-protein interactions directly in single living cells. We previously reported on the detection of homodimer formation in cells using molecular brightness analysis. Here, we extend the technique to detect binding between different proteins. Proteins are labeled with the fluorescent markers YFP and CFP. We first determine the coexpression ratio of both proteins by measuring the intensity ratio with a dual-color setup. The effect of fluorescence resonance energy transfer on the intensity ratio is explicitly taken into account. The brightness of cells coexpressing both proteins is measured in a single-color setup. Selecting the laser wavelength of the two-photon light source allows us to either coexcite both proteins or to selectively excite YFP-labeled proteins. This approach enables us to distinguish between homodimer and heterodimer formation. We first present the theory and then demonstrate experimental feasibility using the ligand binding domains of retinoic acid receptor (RARLBD) and of retinoid X receptor (RXRLBD). Both proteins form heterodimers, and RXRLBD also forms homodimers in the presence of its agonist. We explore binding between these proteins in the presence and absence of RXR agonist. Our results demonstrate that brightness analysis offers a quantitative method for determining protein interactions in cells.
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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.