title: Thermodynamics of peptide binding to the transporter associated with antigen processing (TAP).
abstract: The ATP-binding cassette (ABC) transporter TAP plays an essential role in antigen processing and immune response to infected or malignant cells. TAP translocates proteasomal degradation products from the cytosol into the endoplasmic reticulum, where MHC class I molecules are loaded with these peptides. Kinetically stable peptide-MHC complexes are transported to the cell surface for inspection by cytotoxic T lymphocytes. The transport cycle of TAP is initiated by peptide binding, which is responsible for peptide selection and for stimulation of ATP-hydrolysis and subsequent translocation. Here we have analysed the driving forces for the formation of the peptide-TAP complex by kinetic and thermodynamic methods. First, the apparent peptide association and dissociation rates were determined at various temperatures. Strikingly, very high activation energies for apparent association (E(a)(ass)=106 kJmol(-1)) and dissociation (E(a)(diss)=80 kJmol(-1)) of the peptide-TAP complex were found. Next, the temperature-dependence of the peptide affinity constants was investigated by equilibrium-binding assays. Along with calculations of free enthalpy deltaG, enthalpy deltaH and entropy deltaS, a large positive change in heat capacity was resolved (deltaC degrees =23 kJmol(-1)K(-1)), indicating a fundamental structural reorganization of the TAP complex upon peptide binding. The inspection of the conformational entropy reveals that approximately one-fourth of all TAP residues is rearranged. These thermodynamic studies indicate that at physiological temperature, peptide binding is endothermic and driven by entropy.
authors.affiliation: Institute of Biochemistry, Biocenter, Goethe-University Frankfurt, Marie-Curie-Str. 9, D-60439, Frankfurt, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Neumann
authors.forename: Rupert
authors.initials: R
authors.lastname: Abele
authors.forename: Robert
authors.initials: R
authors.lastname: Tampé
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D017951:Antigen Presentation; D000941:Antigens / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D007700:Kinetics; D010455:Peptides / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D011485:Protein Binding; D013696:Temperature; D013816:Thermodynamics
chemical_list: D018528:ATP-Binding Cassette Transporters; D000941:Antigens; D010455:Peptides