ArticleProceedings of the National Academy of Sciences of the United States of America2010
Heterospecies partition analysis reveals binding curve and stoichiometry of protein interactions in living cells.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- Article
- Identification of the GABARAP binding determinant in PI4K2A.Bioscience reports · 2024Article
- Partitioning of ribonucleoprotein complexes from the cellular actin cortex.Science advances · 2022Article
- Identifying Heteroprotein Complexes in the Nuclear Envelope.Biophysical journal · 2020Article
- Two-Color Spatial Cumulant Analysis Detects Heteromeric Interactions between Membrane Proteins.Biophysical journal · 2019Article
- Fluorescence Imaging Methods to Investigate Translation in Single Cells.Cold Spring Harbor perspectives in biology · 2019Review
- Comobility of GABARAP and Phosphatidylinositol 4-Kinase 2A on Cytoplasmic Vesicles.Biochemistry · 2018Article
- Association of Endophilin B1 with Cytoplasmic Vesicles.Biophysical journal · 2016Article
- Characterization of Ternary Protein Systems In Vivo with Tricolor Heterospecies Partition Analysis.Biophysical journal · 2016Article
- Article
- Quantitative measurement of brightness from living cells in the presence of photodepletion.PloS one · 2014Article
- Time-integrated fluorescence cumulant analysis and its application in living cells.Methods in enzymology · 2013Article
- An elegant way to quantitatively analyze oligomer formation in solution.Biophysical journal · 2012Article
- Molecular brightness analysis reveals phosphatidylinositol 4-Kinase IIβ association with clathrin-coated vesicles in living cells.Biophysical journal · 2012Article
- Fluorescence fluctuation spectroscopy enables quantitative imaging of single mRNAs in living cells.Biophysical journal · 2012Article
- Factors affecting the quantification of biomolecular interactions by fluorescence cross-correlation spectroscopy.Biophysical journal · 2012Article
- Chromophore maturation and fluorescence fluctuation spectroscopy of fluorescent proteins in a cell-free expression system.Analytical biochemistry · 2012Article
- Modern fluorescent proteins and imaging technologies to study gene expression, nuclear localization, and dynamics.Current opinion in cell biology · 2011Review
- Determining antibody stoichiometry using time-integrated fluorescence cumulant analysis.The journal of physical chemistry. B · 2011Article
- Measuring the stoichiometry of functional PspA complexes in living bacterial cells by single molecule photobleaching.Chemical communications (Cambridge, England) · 2011Article
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Abstract
Measuring the binding curve and stoichiometry of protein complexes in living cells is a prerequisite for quantitative modeling of cellular processes. Dual-color fluorescence fluctuation spectroscopy provides a general framework for detecting protein interactions, but lacks suitable methods for quantifying protein heterointeractions in the cell. We address this challenge by introducing heterospecies partition (HSP) analysis for protein heterointeractions of the type D + nA<-->DA(n). HSP directly identifies the heterointeracting species from the sample mixture and determines the binding curve and stoichiometry of the protein complex. The HSP method is applied to provide the first direct characterization of the ligand-dependent binding of the retinoic X receptor to the coactivator transcription intermediate factor 2. A previous study based on protein fragments observed a higher binding stoichiometry than biologically expected. We address this difference in stoichiometry by measuring the binding curves of the full-length proteins in living cells. This study provides proof-of-principle experiments that illustrate the potential of HSP as a general and robust analysis tool for the quantitative characterization of protein heterointeractions by dual-color fluorescence fluctuation spectroscopy in living cells.
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