A kinetics and modeling study of RANTES(9-68) binding to heparin reveals a mechanism of cooperative oligomerization.
Heparan sulfate (HS) and heparin bind to virtually all chemokines and have been shown to play critical roles in the regulation of their activities. However, both binding mechanisms and structural features involved in chemokine-HS interactions remain poorly defined. In the study presented here, we analyzed the binding of heparin to RANTES(9-68), a N-terminally truncated form of the CC-chemokine RANTES. Using biochemical and surface plasmon resonance (BIAcore system) approaches, we showed that the RANTES(9-68)-heparin interaction was characterized by a complex binding model that involved dimerization of the chemokine through a mechanism of positive cooperativity. Since RANTES(9-68) remains monomeric in solution, we concluded that heparin induced chemokine dimerization. The structure of a complex involving a RANTES dimer and a heparin heptadecasaccharide was proposed by molecular modeling. This model was used to design a dimer of "head to head" coupled octasaccharides that would fit the internal symmetry of the chemokine dimer. This engineered oligosaccharide bound RANTES(9-68) much better than a natural heparin fragment of the same length, further supporting the interaction process and the proposed structural model. Altogether, the data reported here provide a basis for understanding the mechanisms by which HS modulates RANTES functions.
Vivès
Romain R
RR
Institut de Biologie Structurale, CNRS-CEA-UJF, 41 rue Horowitz, 38027 Grenoble cedex 01, France.
Sadir
Rabia
R
Imberty
Anne
A
Rencurosi
Anna
A
Lortat-Jacob
Hugues
H
D000818:Animals; D001665:Binding Sites; D015374:Biosensing Techniques; D016466:CHO Cells; D018946:Chemokine CCL5 / Q000737:chemistry* / Q000378:metabolism*; D006224:Cricetinae; D003432:Cross-Linking Reagents / Q000737:chemistry; D019281:Dimerization; D006493:Heparin / Q000737:chemistry* / Q000378:metabolism; D006497:Heparan Sulfate / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D046911:Macromolecular Substances; D008956:Models, Chemical; D008958:Models, Molecular; D009844:Oligosaccharides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D013499:Surface Properties; D013552:Swine
D018946:Chemokine CCL5; D003432:Cross-Linking Reagents; D006493:Heparin; D006497:Heparan Sulfate; D046911:Macromolecular Substances; D009844:Oligosaccharides; C101594:RANTES (9-68)