title: The relationship between ligand-binding thermodynamics and protein-ligand interaction forces measured by atomic force microscopy.
abstract: The interaction forces between biotin and a set of streptavidin site-directed mutants with altered biotin-binding equilibrium and activation thermodynamics have been measured by atomic force microscopy. The AFM technique readily discriminates differences in interaction force between the site-directed (Trp to Phe or Ala) mutants. The interaction force is poorly correlated with both the equilibrium free energy of biotin binding and the activation free energy barrier to dissociation of the biotin-streptavidin complex. The interaction force is generally well correlated with the equilibrium biotin-binding enthalpy as well as the enthalpic activation barrier, but in the one mutant where these two parameters are altered in opposite directions, the interaction force is clearly correlated with the activation enthalpy of dissociation. These results suggest that the AFM force measurements directly probe the enthalpic activation barrier to ligand dissociation.
authors.affiliation: Center for Bioengineering, University of Washington, Seattle 98195, USA.
authors.forename: A
authors.initials: A
authors.lastname: Chilkoti
authors.forename: T
authors.initials: T
authors.lastname: Boland
authors.forename: B D
authors.initials: BD
authors.lastname: Ratner
authors.forename: P S
authors.initials: PS
authors.lastname: Stayton
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D001710:Biotin / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D018625:Microscopy, Atomic Force; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019809:Streptavidin; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D008024:Ligands; D011994:Recombinant Proteins; D001710:Biotin; D019809:Streptavidin