ArticleJournal of the American Chemical Society2009
Detecting protein-ligand binding on supported bilayers by local pH modulation.
Article in Journal of the American Chemical Society, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 59 citations in OpenAlex.
- Lipid-Substrate Interactions Lead to Bilayer Asymmetry.Journal of the American Chemical Society · 2026Article
- Rapid and sensitive detection of cancer-derived small extracellular vesicles using Janus particles.Nature biomedical engineering · 2026Article
- Local pH at Nonionic and Zwitterionic Lipid/Water Interfaces Revealed by Heterodyne-Detected Electronic Sum-Frequency Generation: A Unified View to Predict Interfacial pH of Biomembranes.The journal of physical chemistry. B · 2023Article
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- Engineered molecular sensors for quantifying cell surface crowding.Proceedings of the National Academy of Sciences of the United States of America · 2023Article
- Propranolol induces large-scale remodeling of lipid bilayers: tubules, patches, and holes.RSC advances · 2023Article
- Retromer oligomerization drives SNX-BAR coat assembly and membrane constriction.The EMBO journal · 2023Article
- Characterization of Protein-Phospholipid/Membrane Interactions Using a "Membrane-on-a-Chip" Microfluidic System.Methods in molecular biology (Clifton, N.J.) · 2021Article
- Surface Modification with Control over Ligand Density for the Study of Multivalent Biological Systems.ChemistryOpen · 2020Review
- Self-assembly of l-phenylalanine amino acid: electrostatic induced hindrance of fibril formation.RSC advances · 2019Article
- Topography design in model membranes: Where biology meets physics.Experimental biology and medicine (Maywood, N.J.) · 2019Review
- A Screen for Membrane Fission Catalysts Identifies the ATPase EHD1.Biochemistry · 2019Article
- Dynamin-related protein 1 has membrane constricting and severing abilities sufficient for mitochondrial and peroxisomal fission.Nature communications · 2018Article
- A novel fluorescence microscopic approach to quantitatively analyse protein-induced membrane remodelling.Journal of biosciences · 2018Review
- The RNA-Binding Site of Poliovirus 3C Protein Doubles as a Phosphoinositide-Binding Domain.Structure (London, England : 1993) · 2017Article
- PIP-on-a-chip: A Label-free Study of Protein-phosphoinositide Interactions.Journal of visualized experiments : JoVE · 2017Article
- Calcium Directly Regulates Phosphatidylinositol 4,5-Bisphosphate Headgroup Conformation and Recognition.Journal of the American Chemical Society · 2017Article
- Use of the supported membrane tube assay system for real-time analysis of membrane fission reactions.Nature protocols · 2017Article
- The pleckstrin-homology domain of dynamin is dispensable for membrane constriction and fission.Molecular biology of the cell · 2017Article
- Polyarginine Interacts More Strongly and Cooperatively than Polylysine with Phospholipid Bilayers.The journal of physical chemistry. B · 2016Article
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Herein, we describe a highly sensitive technique for detecting protein-ligand binding at the liquid/solid interface. The method is based upon modulation of the interfacial pH when the protein binds. This change is detected by ortho-Texas Red DHPE, which is doped into supported phospholipid bilayers and used as a pH-sensitive dye. The dye molecule fluoresces strongly at acidic pH values but not basic ones and has an apparent pK(A) of 7.8 in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine membranes containing 0.5 mol % biotin-cap-PE. This method was used to detect antibiotin/biotin binding interactions as well as the binding of cholera toxin B subunits to GM(1). Since these proteins are negatively charged under the conditions of the experiment the interface became slightly more acidic upon binding. In each case, the equilibrium dissociation constant was determined by following the rise in fluorescence as protein was introduced. This change is essentially linear with protein coverage under the conditions employed. For the biotin/antibiotin system it was determined that K(D) = 24 +/- 5 nM, which is in excellent agreement with classical measurements made by total internal reflection fluorescence microscopy involving fluorophore-conjugated antibody molecules. Moreover, the limit of detection was approximately 350 fM at the 99% confidence level. This corresponds to 1 part in 69,000 of the K(D) value. Such a finding compares favorably with surface plasmon resonance studies of similar systems and conditions. The assay could be run in imaging mode to obtain multiple simultaneous measurements using a CCD camera.
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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.