ArticleProceedings of the National Academy of Sciences of the United States of America2017
Weak protein-protein interactions in live cells are quantified by cell-volume modulation.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 67 papers.
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Who cites it
67 citing papers in PubMed.
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- A co-overexpression strategy allows effective protein-protein binding affinities to be assessed as a function of concentration within live cells.bioRxiv : the preprint server for biology · 2026Article
- Too close for comfort: Self-crowding transforms protein structure and stability beyond volume exclusion.Protein science : a publication of the Protein Society · 2026Article
- Proteome-wide prediction of interactions between structured domains and peptide motifs reveals functionally coherent subnetworks.Proceedings of the National Academy of Sciences of the United States of America · 2026Article
- Chemical interactions in polyethylene glycol-induced condensates lead to an anomalous FRET response from a flexible linker-fluorescent protein crowding sensor.bioRxiv : the preprint server for biology · 2026Article
- High Molecular Crowding in Repair Foci Surrounding DNA Breaks, Measured by Fluorescence Lifetime Imaging Microscopy.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2025Article
- Dynamic Cytoplasm: A Physical Regulator of Enzymatic Function.Biochemistry · 2025Review
- Direct Quantification of Protein-Protein Interactions in Living Bacterial Cells.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2025Article
- Direct ionic stress sensing and mitigation by the transcription factor NFAT5.Science advances · 2025Article
- Modulation of Protein-Protein Interactions with Molecular Glues in a Synthetic Condensate Platform.Journal of the American Chemical Society · 2025Article
- Enzymes in a human cytoplasm model organize into submetabolon complexes.Proceedings of the National Academy of Sciences of the United States of America · 2025Article
- Effective in vivo binding energy landscape illustrates kinetic stability of RBPJ-DNA binding.Nature communications · 2025Article
- Probing Electrostatic and Hydrophobic Associative Interactions in Cells.The journal of physical chemistry. B · 2024Article
- A cofactor-induced repressive type of transcription factor condensation can be induced by synthetic peptides to suppress tumorigenesis.The EMBO journal · 2024Article
- Spaces of mathematical chemistry.Theory in biosciences = Theorie in den Biowissenschaften · 2024Article
- Correlating Disordered Activation Domain Ensembles with Gene Expression Levels.bioRxiv : the preprint server for biology · 2024Article
- Hypersensitivity of the vimentin cytoskeleton to net-charge states and Coulomb repulsion.bioRxiv : the preprint server for biology · 2024Article
- Protecting Proteins from Desiccation Stress Using Molecular Glasses and Gels.Chemical reviews · 2024Review
7 more citing papers are in PubMed but not listed here.
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Authors and funding
3 authors.
Funding
Abstract
Weakly bound protein complexes play a crucial role in metabolic, regulatory, and signaling pathways, due in part to the high tunability of their bound and unbound populations. This tunability makes weak binding (micromolar to millimolar dissociation constants) difficult to quantify under biologically relevant conditions. Here, we use rapid perturbation of cell volume to modulate the concentration of weakly bound protein complexes, allowing us to detect their dissociation constant and stoichiometry directly inside the cell. We control cell volume by modulating media osmotic pressure and observe the resulting complex association and dissociation by FRET microscopy. We quantitatively examine the interaction between GAPDH and PGK, two sequential enzymes in the glycolysis catalytic cycle. GAPDH and PGK have been shown to interact weakly, but the interaction has not been quantified in vivo. A quantitative model fits our experimental results with log
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