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Biochemistry. 2009 Sep 22;48(37):8795-805. doi: 10.1021/bi901194p.

Structural basis for differential binding of the interleukin-8 monomer and dimer to the CXCR1 N-domain: role of coupled interactions and dynamics.

Author information

1
Department of Biochemistry and Molecular Biology, Sealy Center for Structural Biology and Molecular Biophysics, 5.142 Medical Research Building, 301 University Boulevard, The University of Texas Medical Branch, Galveston, Texas 77555, USA.

Abstract

Interleukin-8 (IL-8 or CXCL8) plays a critical role in orchestrating the immune response by binding and activating the receptor CXCR1 that belongs to the GPCR class. IL-8 exists as both monomers and dimers, and both bind CXCR1 but with differential affinities. It is well established that the monomer is the high-affinity ligand and that the interactions between the ligand N-loop and receptor N-domain play a critical role in determining binding affinity. In order to characterize the structural basis of differential binding of the IL-8 monomer and dimer to the CXCR1 N-domain, we analyzed binding-induced NMR chemical shift and peak intensity changes and show that they are exquisitely sensitive and can provide detailed insights into the binding process. We used three IL-8 variants, a designed monomer, a trapped disulfide-linked dimer, and WT at dimeric concentrations. NMR data for the monomer show that nonsequential residues that span the entire N-loop are involved in the binding process and that the binding is mediated by a network of extensive direct and indirect coupled interactions. Interestingly, in the case of WT, binding induces dissociation of the dimer-receptor complex to the monomer-receptor complex, and in the case of the trapped dimer, binding results in increased global conformational flexibility. Increased dynamics is evidence of unfavorable interactions, indicating that binding of the WT dimer triggers conformational changes that disrupt dimer-interface interactions, resulting in its dissociation. These results together provide evidence that binding is not a localized event but results in extensive coupled interactions within the monomer and across the dimer interface and that these interactions play a fundamental role in determining binding affinity.

PMID:
19681642
PMCID:
PMC2858980
DOI:
10.1021/bi901194p
[Indexed for MEDLINE]
Free PMC Article

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