TY - JOUR
T1 - Interactions of the Chemokine CCL5/RANTES with Medium-Sized Chondroitin Sulfate Ligands
AU - Deshauer, Courtney
AU - Morgan, Ashli M.
AU - Ryan, Eathen O.
AU - Handel, Tracy M.
AU - Prestegard, James H.
AU - Wang, Xu
N1 - Funding Information:
We would like to thank Dr. Fei Yu of Complex Carbohydrate Research Center for developing the methods for CS dp6 purification, Dr. Brian Cherry of Arizona State University and Dr. John Glushka of the Complex Carbohydrate Research Center for maintaining the NMR spectrometers used in this study, as well as Dr. Andrey Bobkov of Sanford Burnham Medical Research Institute for carrying out ITC experiments. We would also like to thank Connor Zheng, Tsuiying Lan, and Shaghayegh Mazoury for preparing the NMR samples used in the study. We gratefully acknowledge support of the National Institute of General Medical Sciences through an award to X.W. (R00GM088483) and an award to the Resource for Integrated Glycotechnology at the University of Georgia (P41 GM103390), as well as the National Institute for Allergy and Infectious Disease for award RO1-AI37113 to T.M.H. The work also benefitted from instrumentation provided by grant S10 RR027097. The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
Publisher Copyright:
© 2015 Elsevier Ltd.
PY - 2015/6/3
Y1 - 2015/6/3
N2 - Summary Interactions of the chemokine CCL5 (RANTES) with glycosaminoglycans (GAGs) are crucial to the CCL5-mediated inflammation process. However, structural information on interactions between CCL5 and longer GAG fragments is lacking. In this study, the interactions between oligosaccharides derived from chondroitin sulfate and a dimeric variant of CCL5 were investigated using solution nuclear magnetic resonance. The data indicate that, in addition to the BBXB motif in the 40s loop, GAGs also contact residues in the N loop in a manner similar to interactions between chemokine and the receptor N terminus, leading to possible stabilization of the dimer. Using 2,2,6,6-tetramethylpiperidin-1-yl)oxidanyl-tagged hexasaccharides, the binding orientation of the hexasaccharides was shown to be highly dependent on the sulfation pattern of the N-acetyl galactosamine groups. Finally, a model of the CCL5 dimer complexed to chondroitin sulfate hexasaccharides was constructed using paramagnetic relaxation enhancement and intra- and intermolecular nuclear Overhauser effect constraints.
AB - Summary Interactions of the chemokine CCL5 (RANTES) with glycosaminoglycans (GAGs) are crucial to the CCL5-mediated inflammation process. However, structural information on interactions between CCL5 and longer GAG fragments is lacking. In this study, the interactions between oligosaccharides derived from chondroitin sulfate and a dimeric variant of CCL5 were investigated using solution nuclear magnetic resonance. The data indicate that, in addition to the BBXB motif in the 40s loop, GAGs also contact residues in the N loop in a manner similar to interactions between chemokine and the receptor N terminus, leading to possible stabilization of the dimer. Using 2,2,6,6-tetramethylpiperidin-1-yl)oxidanyl-tagged hexasaccharides, the binding orientation of the hexasaccharides was shown to be highly dependent on the sulfation pattern of the N-acetyl galactosamine groups. Finally, a model of the CCL5 dimer complexed to chondroitin sulfate hexasaccharides was constructed using paramagnetic relaxation enhancement and intra- and intermolecular nuclear Overhauser effect constraints.
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U2 - 10.1016/j.str.2015.03.024
DO - 10.1016/j.str.2015.03.024
M3 - Article
C2 - 25982530
AN - SCOPUS:84930384439
SN - 0969-2126
VL - 23
SP - 1066
EP - 1077
JO - Structure
JF - Structure
IS - 6
M1 - 3158
ER -