A universal thermodynamic approach to analyze biomolecular binding experiments.
Binding processes of any kind can be characterized as an association of a given ligand with some binding factor. This includes macromolecules as well as supramolecular aggregates such as micelles or membranes. The underlying molecular binding mechanism may be more or less complicated due to various intermediate steps (involving for instance conformational changes, aggregation, cooperativity, etc.). A sensible discussion of possible binding models naturally calls for a model-independent access to basic thermodynamic properties. The present contribution will demonstrate how this can quite generally be accomplished by a pertinent processing of properly selected experimental data. The method requires a series of titration measurements comprising the use of variable amounts of both the ligand and the binding factor. It leads to a linear mass conservation plot (i.e. amount of the ligand vs. a matching amount of the binding factor) whose slope and ordinate intercept are equal to the binding ratio (i.e. bound ligand per binding factor) and the free ligand concentration, respectively. This establishes the specific binding isotherm. The approach also reveals latent structurally determined features of the applied physical measuring signal. A number of examples including specific binding, unspecific adsorption and insertion in two-dimensional molecular films will illustrate the methodology.
Schwarz
G
G
Department of Biophysical Chemistry, Biocenter of the University of Basel, Switzerland. gerhard.schwarz@unibas.ch
D000408:Alamethicin / Q000378:metabolism; D005456:Fluorescent Dyes; D008024:Ligands; D008081:Liposomes / Q000378:metabolism; D046911:Macromolecular Substances; D008956:Models, Chemical; D009696:Nucleic Acids / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D011479:Protamines / Q000378:metabolism; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D013816:Thermodynamics
D005456:Fluorescent Dyes; D008024:Ligands; D008081:Liposomes; D046911:Macromolecular Substances; D009696:Nucleic Acids; D010455:Peptides; D010743:Phospholipids; D011479:Protamines; D011506:Proteins; D000408:Alamethicin