{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,29]],"date-time":"2026-04-29T11:55:25Z","timestamp":1777463725949,"version":"3.51.4"},"reference-count":84,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2022,8,28]],"date-time":"2022-08-28T00:00:00Z","timestamp":1661644800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Regional Development Fund","award":["UIDB\/04539\/2020"],"award-info":[{"award-number":["UIDB\/04539\/2020"]}]},{"name":"European Regional Development Fund","award":["UIDP\/04539\/2020"],"award-info":[{"award-number":["UIDP\/04539\/2020"]}]},{"name":"European Regional Development Fund","award":["UIDB\/00313\/2020"],"award-info":[{"award-number":["UIDB\/00313\/2020"]}]},{"name":"European Regional Development Fund","award":["UIDP\/00313\/2020"],"award-info":[{"award-number":["UIDP\/00313\/2020"]}]},{"name":"Portuguese funds","award":["UIDB\/04539\/2020"],"award-info":[{"award-number":["UIDB\/04539\/2020"]}]},{"name":"Portuguese funds","award":["UIDP\/04539\/2020"],"award-info":[{"award-number":["UIDP\/04539\/2020"]}]},{"name":"Portuguese funds","award":["UIDB\/00313\/2020"],"award-info":[{"award-number":["UIDB\/00313\/2020"]}]},{"name":"Portuguese funds","award":["UIDP\/00313\/2020"],"award-info":[{"award-number":["UIDP\/00313\/2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The equilibrium distribution of small molecules (ligands) between binding agents in heterogeneous media is an important property that determines their activity. Heterogeneous systems containing proteins and lipid membranes are particularly relevant due to their prevalence in biological systems, and their importance to ligand distribution, which, in turn, is crucial to ligand\u2019s availability and biological activity. In this work, we review several approaches and formalisms for the analysis of the equilibrium distribution of ligands in the presence of proteins, lipid membranes, or both. Special attention is given to common pitfalls in the analysis, with the establishment of the validity limits for the distinct approaches. Due to its widespread use, special attention is given to the characterization of ligand binding through the analysis of Stern\u2013Volmer plots of protein fluorescence quenching. Systems of increasing complexity are considered, from proteins with single to multiple binding sites, from ligands interacting with proteins only to biomembranes containing lipid bilayers and membrane proteins. A new formalism is proposed, in which ligand binding is treated as a partition process, while considering the saturation of protein binding sites. This formalism is particularly useful for the characterization of interaction with membrane proteins.<\/jats:p>","DOI":"10.3390\/ijms23179757","type":"journal-article","created":{"date-parts":[[2022,8,29]],"date-time":"2022-08-29T03:34:56Z","timestamp":1661744096000},"page":"9757","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":26,"title":["Analysis of the Equilibrium Distribution of Ligands in Heterogeneous Media\u2013Approaches and Pitfalls"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3076-9905","authenticated-orcid":false,"given":"Maria Jo\u00e3o","family":"Moreno","sequence":"first","affiliation":[{"name":"Coimbra Chemistry Center\u2014Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"},{"name":"Department of Chemistry, Faculty of Sciences and Technology, University of Coimbra, 3004-535 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1051-2312","authenticated-orcid":false,"given":"Lu\u00eds M. S.","family":"Loura","sequence":"additional","affiliation":[{"name":"Coimbra Chemistry Center\u2014Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"},{"name":"Faculty of Pharmacy, University of Coimbra, 3000-548 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5460-1969","authenticated-orcid":false,"given":"Jorge","family":"Martins","sequence":"additional","affiliation":[{"name":"Centro de Ci\u00eancias do Mar (CCMAR\/CIMAR, LA) and DCBB-FCT, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4442-4900","authenticated-orcid":false,"given":"Armindo","family":"Salvador","sequence":"additional","affiliation":[{"name":"Coimbra Chemistry Center\u2014Institute of Molecular Sciences (CQC-IMS), University of Coimbra, 3004-535 Coimbra, Portugal"},{"name":"CNC\u2014Center for Neuroscience and Cell Biology, 3004-504 Coimbra, Portugal"},{"name":"Institute for Interdisciplinary Research, University of Coimbra, 3030-789 Coimbra, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5702-4538","authenticated-orcid":false,"given":"Adrian","family":"Velazquez-Campoy","sequence":"additional","affiliation":[{"name":"Institute for Biocomputation and Physics of Complex Systems (BIFI), Joint Unit GBsC-CSIC-BIFI, Universidad de Zaragoza, 50018 Zaragoza, Spain"},{"name":"Departamento de Bioqu\u00edmica y Biolog\u00eda Molecular y Celular, Universidad de Zaragoza, 50009 Zaragoza, Spain"},{"name":"Instituto de Investigaci\u00f3n Sanitaria de Arag\u00f3n (IIS Aragon), 50009 Zaragoza, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red en el \u00c1rea Tem\u00e1tica de Enfermedades Hep\u00e1ticas Digestivas (CIBERehd), 28029 Madrid, Spain"}]}],"member":"1968","published-online":{"date-parts":[[2022,8,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/0022-2836(72)90386-5","article-title":"Hydrophobic Free-Energy, Micelle Formation and Association of Proteins with Amphiphiles","volume":"67","author":"Tanford","year":"1972","journal-title":"J. Mol. Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1021\/jp015514e","article-title":"A view of the hydrophobic effect","volume":"106","author":"Southall","year":"2002","journal-title":"J. Phys. Chem. B"},{"key":"ref_3","unstructured":"Creighton, T.E. (1993). Proteins. Structures and Molecular Properties, W. H. Freeman and Company."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"3001","DOI":"10.1016\/S0021-9258(19)41586-X","article-title":"Protein Interactions with Small Molecules\u2014Relationships Between Stoichiometric Binding Constants, Site Binding Constants, and Empirical Binding Parameters","volume":"250","author":"Klotz","year":"1975","journal-title":"J. Biol. Chem."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"011303","DOI":"10.1063\/5.0020997","article-title":"Mechanisms of ligand binding","volume":"1","author":"Cera","year":"2020","journal-title":"Biophys. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Gennis, R.B. (1989). Biomembranes. Molecular Structure and Function, Springer.","DOI":"10.1007\/978-1-4757-2065-5"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1126\/science.175.4023.720","article-title":"Fluid Mosaic Model of Structure of Cell-Membranes","volume":"175","author":"Singer","year":"1972","journal-title":"Science"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"5281","DOI":"10.1021\/bi00414a050","article-title":"Solute Partitioning Into Lipid Bilayer-Membranes","volume":"27","author":"Deyoung","year":"1988","journal-title":"Biochemistry"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1016\/0005-2736(88)90070-3","article-title":"Partitioning of Local-Anesthetics Into Membranes\u2014Surface-Charge Effects Monitored by the Phospholipid Headgroup","volume":"939","author":"Seelig","year":"1988","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/S0005-2736(03)00112-3","article-title":"Quantifying molecular partition into model systems of biomembranes: An emphasis on optical spectroscopic methods","volume":"1612","author":"Santos","year":"2003","journal-title":"Biochim. Biophys. Acta-Biomembr."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7980","DOI":"10.1021\/la0155606","article-title":"On the significance of the solubilization power of detergents","volume":"17","author":"Melo","year":"2001","journal-title":"Langmuir"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"5635","DOI":"10.1021\/j100306a028","article-title":"A Critical Evaluation of Partition Constants in Nonionic Micelles","volume":"91","author":"Melo","year":"1987","journal-title":"J. Phys. Chem."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"562","DOI":"10.1021\/es00003a002","article-title":"Partition of Pesticides of the Coumarin Family between Water and Amphiphilic Aggregates","volume":"29","author":"Lopes","year":"1995","journal-title":"Environ. Sci. Technol."},{"key":"ref_14","unstructured":"Moreno, M.J. (2000). Kinetics of Bimolecular Reactions with Stochastic Effects and Dimensional Constraints, Universidade Nova de Lisboa."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1021\/es00001a017","article-title":"Consequences of the Partition between Water and Molecular Aggregates on the Photodegradation Pattern and Kinetics of a Pesticide of the Coumarin Family","volume":"29","author":"Moreno","year":"1995","journal-title":"Environ. Sci. Technol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"18192","DOI":"10.1021\/jp9614660","article-title":"Stochastic effects on the time-dependent rate constant of photodimerization of 12-(9-anthroyloxy)stearic acid in micelles","volume":"100","author":"Moreno","year":"1996","journal-title":"J. Phys. Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1039\/f29878301391","article-title":"Energy-Transfer in Spherical Geometry\u2014Application to Micelles","volume":"83","author":"Berberansantos","year":"1987","journal-title":"J. Chem. Soc.-Faraday Trans. Ii"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"189","DOI":"10.1016\/S1388-1981(01)00203-7","article-title":"Micelle formation of sodium deoxycholate and sodium ursodeoxycholate (Part 1)","volume":"1580","author":"Matsuoka","year":"2002","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1811","DOI":"10.1073\/pnas.71.5.1811","article-title":"Thermodynamics of Micelle Formation\u2014Prediction of Micelle Size and Size Distribution","volume":"71","author":"Tanford","year":"1974","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Oliver, R.C., Lipfert, J., Fox, D.A., Lo, R.H., Doniach, S., and Columbus, L. (2013). Dependence of Micelle Size and Shape on Detergent Alkyl Chain Length and Head Group. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0062488"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1016\/0927-7757(94)02929-6","article-title":"Factors Determining Emulsion-Type\u2014Hydrophile-Lipophile Balance and Beyond","volume":"91","author":"Davis","year":"1994","journal-title":"Colloids Surf. A"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"121","DOI":"10.1021\/ja00808a020","article-title":"Kinetic Characterization of Bile-Salt Micelles","volume":"96","author":"Menger","year":"1974","journal-title":"J. Am. Chem. Soc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4029","DOI":"10.1021\/j150626a015","article-title":"Measurement of the Rates of Detergent Exchange Between Micelles and the Aqueous Phase Using Phosphorescent Labeled Detergents","volume":"85","author":"Bolt","year":"1981","journal-title":"J. Phys. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/S0001-8686(00)00054-3","article-title":"Solute exchange between surfactant micelles by micelle fragmentation and fusion","volume":"89","author":"Rharbi","year":"2001","journal-title":"Adv. Colloid Interface Sci."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1007\/s00232-017-0009-4","article-title":"Effect of Acyl Chain Length on the Rate of Phospholipid Flip-Flop and Intermembrane Transfer","volume":"251","author":"Vaz","year":"2018","journal-title":"J. Membr. Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"633","DOI":"10.1016\/1074-5521(95)90023-3","article-title":"Lets Get Specific\u2014The Relationship Between Specificity and Affinity","volume":"2","author":"Eaton","year":"1995","journal-title":"Chem. Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"129450","DOI":"10.1016\/j.bbagen.2019.129450","article-title":"Spatial arrangement of LD motif-interacting residues on focal adhesion targeting domain of Focal Adhesion Kinase determine domain-motif interaction affinity and specificity","volume":"1864","author":"Gupta","year":"2020","journal-title":"Biochim. Biophys. Acta-Gen. Subj."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"183157","DOI":"10.1016\/j.bbamem.2019.183157","article-title":"Effect of dipole moment on amphiphile solubility and partition into liquid ordered and liquid disordered phases in lipid bilayers","volume":"1862","author":"Cardoso","year":"2020","journal-title":"Biochim. Biophys. Acta (BBA)-Biomembr."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"3439","DOI":"10.1021\/jp307874v","article-title":"Synthesis and Characterization of a Lipidic Alpha Amino Acid: Solubility and Interaction with Serum Albumin and Lipid Bilayers","volume":"117","author":"Filipe","year":"2013","journal-title":"J. Phys. Chem. B"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"739","DOI":"10.1023\/A:1011923103938","article-title":"Towards the predictability of drug-lipid membrane interactions: The pH-dependent affinity of propranolol to phosphatidylinositol containing liposomes","volume":"15","author":"Kramer","year":"1998","journal-title":"Pharm. Res."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1016\/S0021-9673(00)01266-8","article-title":"Effect of liposome type and membrane fluidity on drug-membrane partitioning analyzed by immobilized liposome chromatography","volume":"913","author":"Liu","year":"2001","journal-title":"J. Chromatogr. A"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.chemphyslip.2008.04.007","article-title":"Partitioning of 1-pyrenesulfonate into zwitterionic and mixed zwitterionic\/anionic fluid phospholipid bilayers","volume":"154","author":"Manuel","year":"2008","journal-title":"Chem. Phys. Lipids"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4184","DOI":"10.1021\/ja209917q","article-title":"Kinetics and Thermodynamics of Chlorpromazine Interaction with Lipid Bilayers: Effect of Charge and Cholesterol","volume":"134","author":"Martins","year":"2012","journal-title":"J. Am. Chem. Soc."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.ab.2009.11.015","article-title":"Partition of amphiphilic molecules to lipid bilayers by isothermal titration calorimetry","volume":"399","author":"Moreno","year":"2010","journal-title":"Anal. Biochem."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1111\/j.1751-1097.1993.tb02314.x","article-title":"Interaction of the Peptide-Hormone Adrenocorticotropin, ACTH(1- 24), with a Membrane Model System\u2014A Fluorescence Study","volume":"57","author":"Moreno","year":"1993","journal-title":"Photochem. Photobiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"3082","DOI":"10.1021\/la703285b","article-title":"NMR study of the sorption behavior of benzyl alcohol derivatives into sonicated and extruded dioctadecyldimethylammonium chloride (DODAC) dispersions: The relevance of membrane fluidity","volume":"24","author":"Saveyn","year":"2008","journal-title":"Langmuir"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1457","DOI":"10.1007\/s11095-007-9263-y","article-title":"Comparing the lipid membrane affinity and permeation of drug-like acids: The intriguing effects of cholesterol and charged lipids","volume":"24","author":"Thomae","year":"2007","journal-title":"Pharm. Res."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1802","DOI":"10.1529\/biophysj.105.060871","article-title":"Permeation of aromatic carboxylic acids across lipid bilayers: The pH-partition hypothesis revisited","volume":"89","author":"Thomae","year":"2005","journal-title":"Biophys. J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"9542","DOI":"10.1128\/JVI.77.17.9542-9552.2003","article-title":"Specific association of glycoprotein B with lipid rafts during herpes simplex virus entry","volume":"77","author":"Bender","year":"2003","journal-title":"J. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"417","DOI":"10.1016\/j.bbalip.2012.10.010","article-title":"The glycolipid transfer protein (GLTP) domain of phosphoinositol 4-phosphate adaptor protein-2 (FAPP2): Structure drives preference for simple neutral glycosphingolipids","volume":"1831","author":"Kamlekar","year":"2013","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"628","DOI":"10.1016\/j.bbamem.2006.12.011","article-title":"Shiga toxin B-subunit sequential binding to its natural receptor in lipid membranes","volume":"1768","author":"Pina","year":"2007","journal-title":"Biochim. Biophys. Acta-Biomembr."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1860","DOI":"10.1016\/j.bbamem.2015.03.014","article-title":"Role of lipid microdomains in TLR-mediated signalling","volume":"1848","author":"Ruysschaert","year":"2015","journal-title":"Biochim. Biophys. Acta-Biomembr."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1016\/S1074-5521(96)90164-7","article-title":"Just add water! The effect of water on the specificity of protein-ligand binding sites and its potential application to drug design","volume":"3","author":"Ladbury","year":"1996","journal-title":"Chem. Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/j.sbi.2010.10.002","article-title":"Protein binding specificity versus promiscuity","volume":"21","author":"Schreiber","year":"2011","journal-title":"Curr. Opin. Struct. Biol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1080\/17460441.2017.1297418","article-title":"A look at ligand binding thermodynamics in drug discovery","volume":"12","author":"Vega","year":"2017","journal-title":"Expert Opin. Drug Discov."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3229","DOI":"10.1007\/s10973-019-08610-0","article-title":"Handling complexity in biological interactions: Allostery and cooperativity in proteins","volume":"138","author":"Vega","year":"2019","journal-title":"J. Therm. Anal. Calorim."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"115","DOI":"10.1016\/j.bpc.2004.12.015","article-title":"ITC in the Post-Genomic era...? Priceless","volume":"115","author":"Freire","year":"2005","journal-title":"Biophys. Chem."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1016\/S0925-4439(97)00027-6","article-title":"Competitive, non-competitive and cooperative interactions between substrates of P-glycoprotein as measured by its ATPase activity","volume":"1361","author":"Litman","year":"1997","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1656","DOI":"10.1021\/acs.molpharmaceut.6b01124","article-title":"Biophysical Study on the Interaction between Eperisone Hydrochloride and Human Serum Albumin Using Spectroscopic, Calorimetric, and Molecular Docking Analyses","volume":"14","author":"Rabbani","year":"2017","journal-title":"Mol. Pharm."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/S0167-4838(01)00287-4","article-title":"Interaction of bovine (BSA) and human (HSA) serum albumins with ionic surfactants: Spectroscopy and modelling","volume":"1594","author":"Gelamo","year":"2002","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1054","DOI":"10.1002\/bio.3071","article-title":"Investigation on the effect of fluorescence quenching of bovine serum albumin by cefoxitin sodium using fluorescence spectroscopy and synchronous fluorescence spectroscopy","volume":"31","author":"Li","year":"2016","journal-title":"Luminescence"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1002\/bip.10489","article-title":"Binding of quercetin with human serum albumin: A critical spectroscopic study","volume":"72","author":"Sengupta","year":"2003","journal-title":"Biopolymers"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"6463","DOI":"10.1021\/ac100721e","article-title":"Development of a Fluorescence Model for the Binding of Medium- to Long-Chain Perfluoroalkyl Acids to Human Serum Albumin Through a Mechanistic Evaluation of Spectroscopic Evidence","volume":"82","author":"Hebert","year":"2010","journal-title":"Anal. Chem."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1007\/s00216-010-3737-1","article-title":"Drug-protein binding: A critical review of analytical tools","volume":"398","author":"Vuignier","year":"2010","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1007\/s00216-003-2111-y","article-title":"Label-free screening of bio-molecular interactions","volume":"377","author":"Cooper","year":"2003","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_56","doi-asserted-by":"crossref","unstructured":"Du, X., Li, Y., Xia, Y.L., Ai, S.M., Liang, J., Sang, P., Ji, X.L., and Liu, S.Q. (2016). Insights into Protein-Ligand Interactions: Mechanisms, Models, and Methods. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17020144"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"2011","DOI":"10.1016\/0006-2952(91)90143-S","article-title":"Analysis of the Binding of Fluorescent Ligands to Soluble- Proteins\u2014Use of Simultaneous Nonlinear Least-Squares Regression to Obtain Estimates of Binding Parameters","volume":"41","author":"Mackay","year":"1991","journal-title":"Biochem. Pharmacol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1016\/S0006-3495(03)74859-0","article-title":"Binding of a Fluorescent Lipid Amphiphile to Albumin and its Transfer to Lipid Bilayer Membranes","volume":"84","author":"Abreu","year":"2003","journal-title":"Biophys. J."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"16337","DOI":"10.1021\/jp105163k","article-title":"Chain Length Effect on the Binding of Amphiphiles to Serum Albumin and to POPC Bilayers","volume":"114","author":"Cardoso","year":"2010","journal-title":"J. Phys. Chem. B"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1023\/A:1009402126863","article-title":"Polarity estimate of the hydrophobic binding sites in erythroid spectrin: A study by pyrene fluorescence","volume":"10","author":"Haque","year":"2000","journal-title":"J. Fluoresc."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Srinivasan, B. (2021). A guide to the Michaelis-Menten equation: Steady state and beyond. FEBS J., 16124.","DOI":"10.1111\/febs.16124"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"1887","DOI":"10.1529\/biophysj.106.086561","article-title":"Exact analysis of heterotropic interactions in proteins: Characterization of cooperative ligand binding by isothermal titration calorimetry","volume":"91","author":"Goni","year":"2006","journal-title":"Biophys. J."},{"key":"ref_63","doi-asserted-by":"crossref","unstructured":"Podjarny, A., Dejaegere, A.P., and Kieffer, B. (2011). Characterisation of Ligand Binding by Calorimetry. Biophysical Approaches Determining Ligand Binding to Biomolecular Targets: Detection, Measurement and Modelling, Royal Society of Chemistry.","DOI":"10.1039\/9781849732666"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"363","DOI":"10.1007\/s00249-021-01509-5","article-title":"Isothermal titration calorimetry (ITC): A standard operating procedure (SOP)","volume":"50","author":"Bastos","year":"2021","journal-title":"Eur. Biophys. J. Biophys. Lett."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.abb.2017.10.020","article-title":"Thermodynamics of cooperative binding of FAD to human NQO1: Implications to understanding cofactor-dependent function and stability of the flavoproteome","volume":"636","author":"Pey","year":"2017","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"185","DOI":"10.1007\/978-1-4939-9179-2_14","article-title":"Tinkering with Binding Polynomials in Isothermal Titration Calorimetry","volume":"Volume 1964","author":"Ennifar","year":"2019","journal-title":"Microcalorimetry of Biological Molecules: Methods and Protocols"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/S0076-6879(08)04205-5","article-title":"Isothermal Titration Calorimetry: General Formalism using Binding Polynomials","volume":"455","author":"Freire","year":"2009","journal-title":"Method. Enzymol."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/j.ymeth.2014.09.010","article-title":"A unified framework based on the binding polynomial for characterizing biological systems by isothermal titration calorimetry","volume":"76","author":"Vega","year":"2015","journal-title":"Methods"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"448","DOI":"10.1016\/j.jmb.2009.08.005","article-title":"A Mechanism for Histone Chaperoning Activity of Nucleoplasmin: Thermodynamic and Structural Models","volume":"393","author":"Taneva","year":"2009","journal-title":"J. Mol. Biol."},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"11891","DOI":"10.1021\/bi00205a027","article-title":"Cooperative Multiple Binding of Bisans and Daunomycin to Tubulin","volume":"33","author":"Ward","year":"1994","journal-title":"Biochemistry"},{"key":"ref_71","unstructured":"Kutner, M., Nachtsheim, C., Neter, J., and Li, W. (2004). Applied Linear Statistical Models, McGraw-Hill\/Irwin. [5th ed.]."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"131979","DOI":"10.1016\/j.chemosphere.2021.131979","article-title":"Critical new insights into the binding of poly- and perfluoroalkyl substances (PFAS) to albumin protein","volume":"287","author":"Alesio","year":"2022","journal-title":"Chemosphere"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"4244","DOI":"10.1529\/biophysj.107.112847","article-title":"Kinetics and thermodynamics of the association of dehydroergosterol with lipid bilayer membranes","volume":"93","author":"Estronca","year":"2007","journal-title":"Biophys. J."},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"319","DOI":"10.1016\/j.colsurfb.2019.04.065","article-title":"Molecular crowding effects on the distribution of amphiphiles in biological media","volume":"180","author":"Martins","year":"2019","journal-title":"Colloids Surf. B"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"1308","DOI":"10.1016\/j.bbamem.2008.02.011","article-title":"Binding of Phospholipids to \u00e1-Lactoglobulin and their Transfer to Lipid Bilayers","volume":"1778","author":"Martins","year":"2008","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1016\/S0167-4838(99)00135-1","article-title":"Enantioselective binding sites on bovine serum albumin to dansyl amino acids","volume":"1433","author":"Abe","year":"1999","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1006\/abio.1995.1290","article-title":"A General, Wide-Range Spectrofluorometric Method for Measuring the Site-Specific Affinities of Drugs Towards Human Serum- Albumin","volume":"227","author":"Epps","year":"1995","journal-title":"Anal. Biochem."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"2423","DOI":"10.1002\/etc.647","article-title":"Strong Associations of Short-Chain Perfluoroalkyl Acids with Serum Albumin and Investigation of Binding Mechanisms","volume":"30","author":"Bischel","year":"2011","journal-title":"Environ. Toxicol. Chem."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1852","DOI":"10.1039\/C9EM00290A","article-title":"Partition coefficients of four perfluoroalkyl acid alternatives between bovine serum albumin (BSA) and water in comparison to ten classical perfluoroalkyl acids","volume":"21","author":"Allendorf","year":"2019","journal-title":"Environ. Sci.-Processes Impacts"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/0005-2736(95)00102-9","article-title":"Partition of Dde in Synthetic and Native Membranes Determined by Ultraviolet Derivative Spectroscopy","volume":"1238","author":"Videira","year":"1995","journal-title":"Biochim. Biophys. Acta-Biomembr."},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"1505","DOI":"10.1023\/B:PHAM.0000036927.37888.93","article-title":"Insertion and partition of sodium taurocholate into egg phosphatidylcholine vesicles","volume":"21","author":"Andrieux","year":"2004","journal-title":"Pharm. Res."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1017\/S0033583508004721","article-title":"Interactions of surfactants with lipid membranes","volume":"41","author":"Heerklotz","year":"2008","journal-title":"Q. Rev. Biophys."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"1914","DOI":"10.1021\/acschemneuro.9b00656","article-title":"Membrane-Dependent Binding and Entry Mechanism of Dopamine into Its Receptor","volume":"11","author":"Lolicato","year":"2020","journal-title":"ACS Chem. Neurosci."},{"key":"ref_84","doi-asserted-by":"crossref","first-page":"1866","DOI":"10.1021\/acs.jpcb.9b10092","article-title":"Menthol Binding to the Human alpha 4 beta 2 Nicotinic Acetylcholine Receptor Facilitated by Its Strong Partitioning in the Membrane","volume":"124","author":"Shahoei","year":"2020","journal-title":"J. Phys. Chem. B"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/17\/9757\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:19:08Z","timestamp":1760141948000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/17\/9757"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,8,28]]},"references-count":84,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["ijms23179757"],"URL":"https:\/\/doi.org\/10.3390\/ijms23179757","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,8,28]]}}}