TY - JOUR
T1 - Oligomeric structure of the chemokine CCL5/RANTES from NMR, MS, and SAXS data
AU - Wang, Xu
AU - Watson, Caroline
AU - Sharp, Joshua S.
AU - Handel, Tracy M.
AU - Prestegard, James H.
N1 - Funding Information:
We thank Bruker AXS for use of their NANOSTAR system and Brian Jones for collecting and evaluating the bioSAXS data on WT CCL5. We thank Dr. Hsiau-Wei Lee for assistance with RDC collection. We also acknowledge the National Center for Research Resources (a part of the NIH) for financial support of the Resource for Integrated Glycotechnology and the CCL5 project (P41-RR005351), the National Institute of General Medical Sciences K99 program for support of X.W. (K99GM088483), and the National Institute of Allergy and Infectious Disease for support of T.M.H. (RO1AI37113 and AI037113-13S1). The content of this work is solely the responsibility of the authors and does not necessarily represent the official views of the NIH.
PY - 2011/8/10
Y1 - 2011/8/10
N2 - CCL5 (RANTES) is a proinflammatory chemokine known to activate leukocytes through its receptor, CCR5. Although the monomeric form of CCL5 is sufficient to cause cell migration in vitro, CCL5's propensity for aggregation is essential for migration in vivo, T cell activation and apoptosis, and HIV entry into cells. However, there is currently no structural information on CCL5 oligomers larger than the canonical CC chemokine dimer. In this study the solution structure of a CCL5 oligomer was investigated using an integrated approach, including NMR residual dipolar couplings to determine allowed relative orientations of the component monomers, SAXS to restrict overall shape, and hydroxyl radical footprinting and NMR cross-saturation experiments to identify interface residues. The resulting model of the CCL5 oligomer provides a basis for explaining the disaggregating effect of E66 and E26 mutations and suggests mechanisms by which glycosaminoglycan binding may promote oligomer formation and facilitate cell migration in vivo.
AB - CCL5 (RANTES) is a proinflammatory chemokine known to activate leukocytes through its receptor, CCR5. Although the monomeric form of CCL5 is sufficient to cause cell migration in vitro, CCL5's propensity for aggregation is essential for migration in vivo, T cell activation and apoptosis, and HIV entry into cells. However, there is currently no structural information on CCL5 oligomers larger than the canonical CC chemokine dimer. In this study the solution structure of a CCL5 oligomer was investigated using an integrated approach, including NMR residual dipolar couplings to determine allowed relative orientations of the component monomers, SAXS to restrict overall shape, and hydroxyl radical footprinting and NMR cross-saturation experiments to identify interface residues. The resulting model of the CCL5 oligomer provides a basis for explaining the disaggregating effect of E66 and E26 mutations and suggests mechanisms by which glycosaminoglycan binding may promote oligomer formation and facilitate cell migration in vivo.
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U2 - 10.1016/j.str.2011.06.001
DO - 10.1016/j.str.2011.06.001
M3 - Article
C2 - 21827949
AN - SCOPUS:80051492024
SN - 0969-2126
VL - 19
SP - 1138
EP - 1148
JO - Structure
JF - Structure
IS - 8
ER -