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The X-Ray Structure of RANTES

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1 The X-Ray Structure of RANTES
Jeffrey P. Shaw, Zoë Johnson, Frédéric Borlat, Catherine Zwahlen, Andreas Kungl, Karen Roulin, Axel Harrenga, Timothy N.C. Wells, Amanda E.I. Proudfoot  Structure  Volume 12, Issue 11, Pages (November 2004) DOI: /j.str

2 Figure 1 Structure of Heparin (A) Heparin polysaccharide structure.
(B) Heparin-derived disaccharide structure. (C) Nomenclature of heparin disaccharides. Structure  , DOI: ( /j.str )

3 Figure 2 Structure of RANTES Dimers
Ribbon diagrams of dimers with space-filling representation of small molecules in the crystal lattice. Red regions in ribbon diagram correspond to regions of the protein where the position of the C-α differs from that of the AOP-RANTES structure by more than 2 Å. A monomers are colored in blue, and B monomers in green. Side chains of residues interacting with small molecules are shown. (A) Dimer of AOP-RANTES with sulfate ions. (B) Dimer of Met-RANTES with sulfate ions. (C) Dimer of RANTES with heparin disaccharide I-S. (D) Dimer of RANTES with heparin disaccharide III-S. (E) Dimer of RANTES-K45E. (F) Dimer of RANTES-44AANA47 with sulfate ions. Structure  , DOI: ( /j.str )

4 Figure 3 Stereo Diagrams of the Binding Site of Heparin Disaccharides in RANTES Crystals and Crystallographic Lattice Heparin disaccharide and residues in close proximity are displayed. Carbon atoms colored in gray are from monomer A, those colored in green or blue are from two different symmetry-related B monomers. Putative hydrogen bonds are shown in violet. (A) Heparin disaccharide I-S with final 2Fo − Fc map contoured at 1σ. (B) Heparin disaccharide III-S with final 2Fo − Fc contoured at 1σ. (C) Crystal lattice of the complex between heparin disaccharide I-S and RANTES. Four dimers of RANTES are displayed. Structure  , DOI: ( /j.str )

5 Figure 4 Structure of RANTES Mutants
The carbon atoms of mutated residues are shown in yellow. (A) Structure of the 40s loop of AOP-RANTES. (B) Structure of the 40s loop of monomer A of RANTES-K45E. (C) Structure of the 40s loop of monomer B of RANTES-K45E. (D) Structure of the 40s loop of monomer C of RANTES-44AANA47. (E) Structure of 40s loop of monomer D of RANTES-44AANA47. (F) Structure of RANTES-44AANA47 near the sulfate ion binding to the 50s loop. Final 2Fo-Fc contoured at 1σ. Structure  , DOI: ( /j.str )

6 Figure 5 Biochemical Characterization and In Vivo Behavior of RANTES Mutants (A) Relative affinity of mutant and wild-type RANTES for heparin octasaccharide. Black squares, wild-type RANTES; inverted white triangles, RANTES-44AANA47; black triangles, RANTES-K45E; and white triangles, RANTES-Y3A for a heparin octasaccharide as measured by isothermal fluorescence titrations. (B) Inhibition by RANTES-Y3A of peritoneal cellular recruitment induced by RANTES. Ten micrograms of RANTES-Y3A is unable to induce peritoneal recruitment (light gray bar). The peritoneal cell recruitment induced by 10 μg of RANTES can be inhibited by RANTES-Y3A in a dose-dependant manner (gray bars). (C) Inhibition by RANTES-K45E of peritoneal cellular recruitment induced by RANTES. In a similar manner, RANTES-K45E (light gray bars) is unable to induce peritoneal cell recruitment. The peritoneal cell recruitment induced by RANTES (black bar) is also inhibited in a dose-dependent manner by RANTES-K45E (gray bars). Structure  , DOI: ( /j.str )

7 Figure 6 Inhibition of Peritoneal Cellular Recruitment Induced by RANTES by Heparin-Derived Oligosaccharides The peritoneal cell recruitment induced by 10 μg of RANTES (black bar) can be inhibited by pretreatment, 30 min prior to RANTES injection, by injection of 20 μg of heparin (stippled bar). Pretreatment with a heparin-derived disaccharide at a 15-fold molar excess, compared to the injected RANTES, has no effect on RANTES-mediated cell recruitment. The inhibitory effect of pretreatment with heparin-derived tetra-, hexa- and octasaccharides depends on the molar excess of oligosaccharides. Structure  , DOI: ( /j.str )


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