Solution NMR characterization of chemokine CXCL8/IL-8 monomer and dimer binding to glycosaminoglycans: structural plasticity mediates differential binding interactions

Joseph, Prem Raj B.; Mosier, Philip D.; Desai, Umesh R.; Rajarathnam, Krishna

doi:10.1042/bj20150059

Cited by 95 publications

(118 citation statements)

References 66 publications

(71 reference statements)

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“…This suggests that chemical shift changes from direct and indirect interactions are of opposite sign and similar magnitude and, therefore, cancel out. We made similar observations earlier for CXCL8, where lysines known to be involved in GAG binding from functional studies showed minimal backbone chemical shift changes (26).…”

Section: Characterization Of Gag-hcxcl1supporting

confidence: 66%

“…Interestingly, CXCL5, CXCL8, and KC actually have an acidic residue at the third ␤-strand position. NMR studies of CXCL8 and KC show evidence of binding only to the ␣-domain (26,32). In addition, only CXCL1, CXCL2, and CXCL3 have a basic residue at the end of the first ␤-strand.…”

Section: Discussionmentioning

confidence: 99%

“…We have shown recently that heparin-binding residues in CXCL8 can be classified as core and peripheral residues and that GAG can bind in different geometries (26). The core residues in CXCL8 are located in the N-loop and C-terminal helix, and these residues are also involved in hCXCL1-GAG interactions.…”

Section: Discussionmentioning

confidence: 99%

“…Docking of hCXCL1-Heparin Complexes-Molecular docking of heparin oligosaccharides to hCXCL1 WT and the R8A dimer was carried out using high ambiguity driven biomolecular docking (HADDOCK) (24,25), as described previously for the CXCL8-heparin complexes (26). We used the hCXCL1 dimer (PDB code 1MGS) and dp8-and dp14-mer structures generated from a heparin 12-mer (PDB code 1HPN) as the starting structures (19,27).…”

Section: Methodsmentioning

confidence: 99%

“…Considering that buried residues cannot be involved in direct binding, their perturbations must be due to indirect interactions, such as binding-induced structure formation or structural rearrangement. For instance, CSPs of the helical residues Ile-58 and Ile-62 must be due to rearrangement of the ␣-helix, and a similar rearrangement of the helix on GAG binding to CXCL8 has also been observed (26).…”

Section: Characterization Of Gag-hcxcl1mentioning

confidence: 99%

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CXCL1/MGSA Is a Novel Glycosaminoglycan (GAG)-binding Chemokine

Sepuru

Rajarathnam

2016

Journal of Biological Chemistry

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In humans, the chemokine CXCL1/MGSA (hCXCL1) plays fundamental and diverse roles in pathophysiology, from microbial killing to cancer progression, by orchestrating the directed migration of immune and non-immune cells. Cellular trafficking is highly regulated and requires concentration gradients that are achieved by interactions with sulfated glycosaminoglycans (GAGs). However, very little is known regarding the structural basis underlying hCXCL1-GAG interactions. We addressed this by characterizing the binding of GAG heparin oligosaccharides to hCXCL1 using NMR spectroscopy. Binding experiments under conditions at which hCXCL1 exists as monomers and dimers indicate that the dimer is the high-affinity GAG ligand. NMR experiments and modeling studies indicate that lysine and arginine residues mediate binding and that they are located in two non-overlapping domains. One domain, consisting of N-loop and C-helical residues (defined as ␣-domain) has also been identified previously as the GAG-binding domain for the related chemokine CXCL8/IL-8. The second domain, consisting of residues from the N terminus, 40s turn, and third ␤-strand (defined as ␤-domain) is novel. Eliminating ␤-domain binding by mutagenesis does not perturb ␣-domain binding, indicating two independent GAG-binding sites. It is known that N-loop and N-terminal residues mediate receptor activation, and we show that these residues are also involved in extensive GAG interactions. We also show that the GAG-bound hCXCL1 completely occlude receptor binding. We conclude that hCXCL1-GAG interactions provide stringent control over regulating chemokine levels and receptor accessibility and activation, and that chemotactic gradients mediate cellular trafficking to the target site.Chemokines, a large family of small soluble proteins, are highly versatile and play fundamental roles in diverse functions, from combating infection and initiating tissue repair to regulating metabolism and organ development (1, 2). Common to these various functions is the directed movement of various cell types to distal and remote locations. Cellular trafficking must be highly coordinated to elicit the required biological function, and its dysregulation could be detrimental, resulting in disease. There is now increasing evidence that the ability of a chemokine to reversibly exist as monomers and dimers and binding glycosaminoglycans is coupled and plays important roles in mediating these functions (3, 4).Glycosaminoglycans (GAGs) 2 such as heparan sulfate (HS) are highly sulfated polysaccharides. They are expressed ubiquitously by many cell types, are anchored to the cell surface by covalent attachment to membrane proteins, and form non-covalent complexes with proteins in the extracellular matrix (5-7). Animal models and cellular studies have established that GAG interactions dictate chemokine concentration gradients and that these gradients orchestrate cellular trafficking (8 -10). GAGs are acidic, and chemokines are basic or contain clusters of basic residues, indicating that electr...

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Section: Characterization Of Gag-hcxcl1supporting

confidence: 66%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Characterization Of Gag-hcxcl1mentioning

confidence: 99%

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CXCL1/MGSA Is a Novel Glycosaminoglycan (GAG)-binding Chemokine

Sepuru

Rajarathnam

2016

Journal of Biological Chemistry

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Chemoenzymatic Synthesis of Heparan Sulfate Oligosaccharides having a Domain Structure**

2022

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Heparan sulfate (HS) has a domain structure in which regions that are modified by epimerization and sulfonation (NS domains) are interspersed by unmodified fragments (NA domains). There is data to support that domain organization of HS can regulate binding of proteins, however, such model has been difficult to probe. Here, we report a chemoenzymatic methodology that can provide HS oligosaccharides composed of two or more NS domains separated by NA domains of different length. It is based on the chemical synthesis of a HS oligosaccharide that enzymatically was extended by various GlcA-GlcNAc units and terminated in GlcNAc having an azido moiety at C-6 position. HS oligosaccharides having an azide and alkyne moiety could be assembled by copper catalyzed alkyne-azide cycloaddition to give compounds having various NS domains separated by unsulfonated regions. Competition binding studies showed that the length of an NA domain modulates the binding of the chemokines CCL5 and CXCL8.

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Chemoenzymatic Synthesis of Heparan Sulfate Oligosaccharides having a Domain Structure**

2022

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Heparan sulfate (HS) have domain structures in which regions that are substantially modified by epimerization and sulfation (NS domains) are interspersed by unmodified fragments (NA domains). There is data to support that the domain structure of HS can regulate protein binding, however, such a binding mode has been difficult to probe. Here, we report a chemoenzymatic methodology that can provide HS oligosaccharides composed of two or more NS domains separated by NA domains of different length. It is based on the chemical synthesis of a sulfated HS oligosaccharide that enzymatically could be extended by various GlcA-GlcNAc units and terminated in a GlcNAc-6N3 moiety. HS oligosaccharides having an azide and alkyne moiety could assembled by copper catalyzed alkyne-azide cycloaddition (CuAAC) to give compounds having various NS domains separated by unsulfated regions. Competition binding studies showed that the length of an NA domain modulates the binding of the chemokines CCL5 and CXCL8.

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Solution NMR characterization of chemokine CXCL8/IL-8 monomer and dimer binding to glycosaminoglycans: structural plasticity mediates differential binding interactions

Cited by 95 publications

References 66 publications

CXCL1/MGSA Is a Novel Glycosaminoglycan (GAG)-binding Chemokine

CXCL1/MGSA Is a Novel Glycosaminoglycan (GAG)-binding Chemokine

Chemoenzymatic Synthesis of Heparan Sulfate Oligosaccharides having a Domain Structure**

Chemoenzymatic Synthesis of Heparan Sulfate Oligosaccharides having a Domain Structure**

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