Structure of a CXC chemokine-receptor fragment in complex with interleukin-8

Skelton, Nicholas J.; Quan, Cliff; Reilly, Dorothea; Lowman, Henry B.

doi:10.1016/s0969-2126(99)80022-7

Cited by 166 publications

(253 citation statements)

References 57 publications

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“…Residues in the N-terminal domain (binding site-I) and in extracellular loops (binding site-II) of CXCR1 interact with IL-8 (13,14,16,(30)(31)(32)(33)35,66,68,69). Specific residues in the N-terminal domain of CXCR1 contribute to both the selectivity and the affinity of the receptor for IL-8.…”

Section: Discussionmentioning

confidence: 99%

“…The IL-8 residues that directly interact with various N-terminal CXCR1 constructs, including ND-CXCR1 (1-38) and 1TM-CXCR1 (1-72), are well defined (15,30). The residues in the N-loop and 40s loop of IL-8 contribute to its major binding sites and display some evidence of local motions (14,15,31,33). In combination with the solid-state NMR data obtained on IL-8 (1-66) bound to CXCR1 (1-350) and NT-CXCR1 (39-350) in phospholipid bilayers, this suggests that the dynamics of free and bound IL-8 may play important roles in receptor binding selectivity as well as activation.…”

Section: Discussionmentioning

confidence: 99%

“…1 B. Although none of the structures represent complexes of a receptor with one of its native chemokine agonists, the structures of receptors bound to viral chemokines provide information that is complementary to the many mutagenesis and binding studies of their interactions. There have been many solution NMR and other biophysical studies of IL-8 interacting with polypeptides whose sequences correspond to N-terminal residues of CXCR1 (13)(14)(15)(16)(30)(31)(32)(33). The current model for chemokine-receptor interactions involves two binding sites on the receptor.…”

Section: Introductionmentioning

confidence: 99%

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Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Park

Berkamp

Radoicic

et al. 2017

Biophysical Journal

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The human chemokine interleukin-8 (IL-8; CXCL8) is a key mediator of innate immune and inflammatory responses. This small, soluble protein triggers a host of biological effects upon binding and activating CXCR1, a G proteincoupled receptor, located in the cell membrane of neutrophils. Here, we describe 1 H-detected magic angle spinning solid-state NMR studies of monomeric IL-8 (1-66) bound to full-length and truncated constructs of CXCR1 in phospholipid bilayers under physiological conditions. Cross-polarization experiments demonstrate that most backbone amide sites of IL-8 (1-66) are immobilized and that their chemical shifts are perturbed upon binding to CXCR1, demonstrating that the dynamics and environments of chemokine residues are affected by interactions with the chemokine receptor. Comparisons of spectra of IL-8 (1-66) bound to full-length CXCR1 (1-350) and to N-terminal truncated construct NT-CXCR1 (39-350) identify specific chemokine residues involved in interactions with binding sites associated with N-terminal residues (binding site-I) and extracellular loop and helical residues (binding site-II) of the receptor. Intermolecular paramagnetic relaxation enhancement broadening of IL-8 (1-66) signals results from interactions of the chemokine with CXCR1 (1-350) containing Mn 2þ chelated to an unnatural amino acid assists in the characterization of the receptor-bound form of the chemokine.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Park

Berkamp

Radoicic

et al. 2017

Biophysical Journal

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“…1,2 CXC chemokines are small proteins (8)(9)(10)(11)(12) kDa) that contain a conserved CXC residue motif (C-cysteine, X-any other residue) proximal to the N-terminal region of the protein. 3 Solution and solid-state structures of several human CXC chemokines have been solved, revealing a common tertiary fold for all members of the family consisting of an unstructured N-terminal loop, antiparellel b-strands and a C-terminal a-helix.…”

Section: Introductionmentioning

confidence: 99%

“…The dissociation constant (K d ) for CXCL8 dimerization has been reported to be 10 to 20 lM. 7,9 As CXCL8 is secreted at high concentrations from injured or cancerous tissues, its local concentration can vary significantly, leading to the existence of both the monomeric and dimeric forms at different locations and over time. 10,11 This differential distribution of the CXCL8 dimeric and monomeric forms may suggest an important role for each of them in the functional activity of this chemokine.…”

Section: Introductionmentioning

confidence: 99%

The dynamics of interleukin‐8 and its interaction with human CXC receptor I peptide

et al. 2014

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Interleukin-8 (CXCL8, IL-8) is a proinflammatory chemokine important for the regulation of inflammatory and immune responses via its interaction with G-protein coupled receptors, including CXC receptor 1 (CXCR1). CXCL8 exists as both a monomer and as a dimer at physiological concentrations, yet the molecular basis of CXCL8 interaction with its receptor as well as the importance of CXCL8 dimer formation remain poorly characterized. Although several biological studies have indicated that both the CXCL8 monomer and dimer are active, biophysical studies have reported conflicting results regarding the binding of CXCL8 to CXCR1. To clarify this problem, we expressed and purified a peptide (hCXCR1pep) corresponding to the N-terminal region of human CXCR1 (hCXCR1) and utilized nuclear magnetic resonance (NMR) spectroscopy to interrogate the binding of wild-type CXCL8 and a previously reported mutant (CXCL8M) that stabilizes the monomeric form. Our data reveal that the CXCL8 monomer engages hCXCR1pep with a slightly higher affinity than the CXCL8 dimer, but that the CXCL8 dimer does not dissociate upon binding hCXCR1pep. These investigations also showed that CXCL8 is dynamic on multiple timescales, which may help explain the versatility in this interleukin for engaging its target receptors.

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Receptor-binding conformation of the ?ELR? motif of IL-8: X-ray structure of the L5C/H33C variant at 2.35 ? resolution

Gerber¹,

Lowman²,

Artis³

2000

Proteins

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The "ELR" (Glu-Leu-Arg) tripeptide sequence near the N-terminus of interleukin-8 (IL-8) contributes a large part of the receptor binding free energy. Prior X-ray and nuclear magnetic resonance (NMR) structures of IL-8 have shown this region of the molecule to be highly mobile. We reasoned that a hydrophobic interaction between the leucine and the neighboring beta-turn might exist in the receptor binding conformation of the N-terminus. To test this hypothesis, we mutated two residues to cysteine and connected the N-terminus to the beta-turn. The mutant retains receptor binding affinity reasonably close to wild type and allows the characterization of a high-affinity conformation that may be useful in the design of small IL-8 mimics. The L5C/H33C mutant is refined to R-values of R = 20.6% and Rfree = 27.7% at 2.35 A resolution. Other receptor binding determinants reside in the "N-loop" found after "ELR" and preceding the first beta-strand. All available structures of IL-8 have been found with one of two distinct N-loop conformations. One of these is relevant for receptor binding, based on NMR results with receptor peptides. The other conformation obscures the receptor-peptide binding surface and may have an undetermined but necessarily different function.

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Structure of a CXC chemokine-receptor fragment in complex with interleukin-8

Cited by 166 publications

References 57 publications

Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

Interaction of Monomeric Interleukin-8 with CXCR1 Mapped by Proton-Detected Fast MAS Solid-State NMR

The dynamics of interleukin‐8 and its interaction with human CXC receptor I peptide

Receptor-binding conformation of the ?ELR? motif of IL-8: X-ray structure of the L5C/H33C variant at 2.35 ? resolution

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