title: Insights into the mechanism by which interferon-γ basic amino acid clusters mediate protein binding to heparan sulfate.
abstract: The extensive functional repertoire of heparin and heparan sulfate, which relies on their ability to interact with a large number of proteins, has recently emerged. To understand the forces that drive such interactions the binding of heparin to interferon-γ (IFNγ), used as a model system, was investigated. NMR-based titration experiments demonstrated the involvement of two adjacent cationic domains (D1: KTGKRKR and D2: RGRR), both of which are present within the carboxy-terminal sequence of the cytokine. Kinetic analysis showed that these two domains contribute differently to the interaction: D1 is required to form a complex and constitutes the actual binding site, whereas D2, although unable to associate with heparin by itself, increased the association rate of the binding. These data are consistent with the view that D2, through nonspecific electrostatic forces, places the two molecules in favorable orientations for productive binding within the encounter complex. This mechanism was supported by electrostatic potential analysis and thermodynamic investigations. They showed that D1 association to heparin is driven by both favorable enthalpic and entropic contributions, as expected for a binding sequence, but that D2 gives rise to entropic penalty, which opposes binding in a thermodynamic sense. The binding mechanism described herein, by which the D2 domain kinetically drives the interaction, has important functional consequences and gives a structural framework to better understand how specific are the interactions between proteins and heparin.
authors.affiliation: Institut de Biologie Structurale, CNRS, CEA, University Grenoble Alpes, UMR 5075, 41 rue Horowitz, 38027, Grenoble cedex 01, France.
authors.forename: Els
authors.initials: E
authors.lastname: Saesen
authors.forename: Stéphane
authors.initials: S
authors.lastname: Sarrazin
authors.forename: Cédric
authors.initials: C
authors.lastname: Laguri
authors.forename: Rabia
authors.initials: R
authors.lastname: Sadir
authors.forename: Damien
authors.initials: D
authors.lastname: Maurin
authors.forename: Aline
authors.initials: A
authors.lastname: Thomas
authors.forename: Anne
authors.initials: A
authors.lastname: Imberty
authors.forename: Hugues
authors.initials: H
authors.lastname: Lortat-Jacob
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D006497:Heparan Sulfate / Q000737:chemistry*; D007371:Interferon-gamma / Q000737:chemistry* / Q000235:genetics; D009154:Mutation; D011506:Proteins / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D006497:Heparan Sulfate; D007371:Interferon-gamma; D011506:Proteins