The reported structures of
many CC chemokines show a conserved dimer interface along theirN-terminal region, raising the possibility that the quaternary arrangement of these s
mall immune proteinsmight influence their function. We have produced and analyzed several mutants of MIP-1
![](/i<font color=)
mages/gifchars/beta2.gif" BORDER=0 ALIGN="middle"> having arange of dimer
Kd values in order to determine the significance of dimerization in receptor binding andcellular activation. NMR and analytical ultracentrifugation were used to analyze the oligomeric state ofthe mutants. Functional relevance was determined by receptor binding affinity and the ability to invokeintracellular calcium release from CHO cells transfected with the MIP-1
![](/i<font color=)
mages/gifchars/beta2.gif" BORDER=0 ALIGN="middle"> receptor CCR5. The monomericN-terminally truncated mutant MIP(9) was able to bind the CCR5 receptor with a
Ki of 600 pM butdisplayed weak agonistic properties, while the monomeric mutant P8A still retained the ability to tightlybind (
Ki = 480 pM) and to activate (EC
50 = 12 nM) the receptor. These data suggest that the MIP-1
![](/i<font color=)
mages/gifchars/beta2.gif" BORDER=0 ALIGN="middle">dimer is not required for CCR5 binding or activation. In addition, we identified Phe13, the residueimmediately following the conserved CC motif in MIP-1
![](/i<font color=)
mages/gifchars/beta2.gif" BORDER=0 ALIGN="middle">, as a key determinant for binding to CCR5.Replacement of Phe13 by Tyr, Leu, Lys, and Ala showed the aro
matic side chain to be important for bothbinding to CCR5 and chemokine dimerization.