Backbone dynamics of the human CC-chemokine eotaxin.
Eotaxin is a CC chemokine with potent chemoattractant activity towards eosinophils. 15N NMR relaxation data have been used to characterize the backbone dynamics of recombinant human cotaxin. 15N longitudinal (R1) and transverse (R2) auto relaxation rates, heteronuclear ¿1H¿-15N steady-state NOEs, and transverse cross-relaxation rates (eta xy) were obtained at 30 degrees C for all resolved backbone secondary amide groups using 1H-detected two-dimensional NMR experiments. Ratios of transverse auto and cross relaxation rates were used to identify NH groups influenced by slow conformational rearrangement. Relaxation data were fit to the extended model free dynamics formalism, yielding parameters describing axially symmetric molecular rotational diffusion and the internal dynamics of each NH group. The molecular rotational correlation time (tau m) is 5.09 +/- 0.02 ns, indicating that eotaxin exists predominantly as a monomer under the conditions of the NMR study. The ratio of diffusion rates about unique and perpendicular axes (D parallel/D perpendicular) is 0.81 +/- 0.02. Residues with large amplitudes of subnanosecond motion are clustered in the N-terminal region (residues 1-19), the C-terminus (residues 68-73) and the loop connecting the first two beta-strands (residues 30-37). N-terminal flexibility appears to be conserved throughout the chemokine family and may have implications for the mechanism of chemokine receptor activation. Residues exhibiting significant dynamics on the microsecond-millisecond time scale are located close to the two conserved disulfide bonds, suggesting that these motions may be coupled to disulfide bond isomerization.
Ye
J
J
Department of Chemistry, Indiana University, Bloomington 47405-0001, USA.
Mayer
K L
KL
Stone
M J
MJ
D016880:Anisotropy; D054413:Chemokine CCL11; D018925:Chemokines / Q000737:chemistry; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil / Q000737:chemistry; D016207:Cytokines / Q000737:chemistry*; D004220:Disulfides; D006801:Humans; D007536:Isomerism; D008432:Mathematical Computing; D008958:Models, Molecular; D009038:Motion; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017433:Protein Structure, Secondary; D012399:Rotation
C515456:CCL11 protein, human; D054413:Chemokine CCL11; D018925:Chemokines; D019742:Chemokines, CC; D002631:Chemotactic Factors, Eosinophil; D016207:Cytokines; D004220:Disulfides