Journal: The Journal of Biological Chemistry

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

doi: 10.1074/jbc.M115.697888

Figure Lengend Snippet: The hCXCL1 dimer is the high-affinity GAG ligand. A section of the 1H,15N HSQC spectra showing the overlay of WT hCXCL1 in the free (black) and GAG-bound (red) forms. Dimer (d) and monomer (m) peaks are indicated. The monomer peaks disappear on dp14 binding, indicating that the dimer is the high-affinity GAG ligand. The spectra were collected using a 15 μm hCXCL1 sample in 50 mm sodium phosphate (pH 6.0) at 40 °C.

Article Snippet: The heparin oligosaccharides dp8 and dp14 were purchased from Iduron.

Techniques: Binding Assay

Journal: The Journal of Biological Chemistry

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

doi: 10.1074/jbc.M115.697888

Figure Lengend Snippet: Binding of WT hCXCL1 to heparin GAGs. A, sections of the 1H,15N HSQC spectra showing the overlay of WT hCXCL1 in the free (black) and dp14-bound (red) forms. Arrows indicate the direction of the peak movement. B and C, histograms of chemical shift changes in the hCXCL1 dimer on binding heparin dp8 (B) and dp14 (C). The basic residues Lys, Arg, and His are shown in blue, and buried residues (ASA < 20%) are shown in black. The CSP of Lys-21 is truncated, and the actual CSP is 1.80 ppm. The horizontal line at 0.1 ppm represents the cutoff for a residue to be considered perturbed. The spectra were collected using a 100 μm hCXCL1 sample in 50 mm sodium phosphate (pH 5.7) at 40 °C.

Article Snippet: The heparin oligosaccharides dp8 and dp14 were purchased from Iduron.

Techniques: Binding Assay

Journal: The Journal of Biological Chemistry

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

doi: 10.1074/jbc.M115.697888

Figure Lengend Snippet: Backbone dynamics of the hCXCL1-GAG complex. Comparison of 15N,1H NOE values for hCXCL1 in the free (black) and dp14-bound (red) forms. The data show that the N-terminal, N-loop, and 30s loop residues in the GAG-bound form have higher NOE values. A NOE difference plot between the bound and free forms (NOEbound − NOEfree) is shown as an inset. The spectra were collected using a 100 μm hCXCL1 sample in 50 mm sodium phosphate (pH 5.7) at 40 °C.

Article Snippet: The heparin oligosaccharides dp8 and dp14 were purchased from Iduron.

Techniques:

Journal: The Journal of Biological Chemistry

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

doi: 10.1074/jbc.M115.697888

Figure Lengend Snippet: Binding of the hCXCL1 R8A mutant to heparin GAG. Shown is a histogram of chemical shift changes in the hCXCL1 R8A mutant on binding heparin dp14. The data indicate binding only to the α-domain. The basic residues Lys, Arg, and His are shown in blue, and buried residues (ASA < 20%) are shown in black. The CSP of Lys-21 is truncated, and the actual CSP is 1.85 ppm. The horizontal line at 0.1 ppm represents the cutoff for a residue to be considered perturbed. The spectra were collected using a 100 μm hCXCL1 sample in 50 mm sodium phosphate (pH 5.7) at 40 °C.

Article Snippet: The heparin oligosaccharides dp8 and dp14 were purchased from Iduron.

Techniques: Binding Assay, Mutagenesis