title: Dimer dissociation is essential for interleukin-8 (IL-8) binding to CXCR1 receptor.
abstract: Chemokines play a fundamental role in trafficking of immune cells and in host defense against infection. The role of chemokines in the recruitment process is highly regulated spatially and temporally and involves interactions with G protein-coupled receptors and cell surface glycosaminoglycans. The dynamic equilibrium between chemokine monomers and dimers, both free in solution and in cell surface-bound forms, regulates different components of recruitment such as chemotaxis and receptor signaling. The binding and activity of the chemokine interleukin-8 (IL-8) for its receptors, previously studied using "trapped" non-associating monomers and non-dissociating dimers, show that the monomer has a native-like function but support conflicting roles for the dimer. We have measured the binding of native IL-8 to the CXCR1 N-domain, using isothermal titration calorimetry and sedimentation equilibrium techniques. The N-domain constitutes a critical binding site, and IL-8 binding affinity to the receptor N-domain is in the same concentration range as the IL-8 monomerdimer equilibrium. We observed that only the IL-8 monomer, and not the dimer, is competent in binding the receptor N-domain. Based on our results, we propose that IL-8 dimerization functions as a negative regulator for the receptor function and as a positive regulator for binding to glycosaminoglycans and that both play a role in the neutrophil recruitment process.
authors.affiliation: Department of Human Biological Chemistry and Genetics, and Sealy Center for Structural Biology, University of Texas Medical Branch, Galveston, TX 77555-1055, USA.
authors.forename: Harshica
authors.initials: H
authors.lastname: Fernando
authors.forename: Christopher
authors.initials: C
authors.lastname: Chin
authors.forename: Jörg
authors.initials: J
authors.lastname: Rösgen
authors.forename: Krishna
authors.initials: K
authors.lastname: Rajarathnam
mesh_terms: D000818:Animals; D001665:Binding Sites; D002151:Calorimetry; D002851:Chromatography, High Pressure Liquid; D019281:Dimerization; D006025:Glycosaminoglycans / Q000737:chemistry; D016209:Interleukin-8 / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008956:Models, Chemical; D009504:Neutrophils / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011817:Rabbits; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry; D023062:Receptors, Interleukin-8A / Q000737:chemistry / Q000378:metabolism*; D013816:Thermodynamics; D014461:Ultracentrifugation
chemical_list: D006025:Glycosaminoglycans; D016209:Interleukin-8; D008024:Ligands; D010455:Peptides; D043562:Receptors, G-Protein-Coupled; D023062:Receptors, Interleukin-8A