PMID- 15544331
OWN - NLM
STAT- MEDLINE
DCOM- 20050118
LR  - 20071115
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 43
IP  - 46
DP  - 2004 Nov 23
TI  - Chemokine-binding specificity of soluble chemokine-receptor analogues: 
      identification of interacting elements by chimera complementation.
PG  - 14602-11
AB  - The specificity of chemokine-receptor interactions plays a central role in the 
      regulation of leukocyte migration in inflammatory responses. Herein, we describe 
      a soluble mimic of CC chemokine receptor 2 (CCR2), dubbed CROSS-N(2)E3(2), which 
      incorporates the N-terminal region (N) and third extracellular loop (E3) elements 
      of CCR2 displayed on the surface of a soluble protein scaffold. CROSS-N(2)E3(2) 
      binds to the CCR2 ligand monocyte chemoattractant protein-1 (MCP-1) with a 
      dissociation equilibrium constant of 1.1 +/- 0.1 microM but does not bind to the 
      cognate chemokines of the receptor CCR3 (eotaxin-1, -2, and -3). Similarly, a 
      soluble analogue of CCR3 (CROSS(5)-N(3)E3(3)) binds to eotaxin-1, -2, and -3 but 
      not to MCP-1. Thus, these receptor analogues have the same specificity as the 
      natural receptors. Using soluble proteins containing N and E3 elements from 
      different receptors (CROSS-N(2)E3(3) and CROSS-N(3)E3(2)), we demonstrate that 
      both receptor elements are required for optimal binding to the cognate 
      chemokines. In addition, we report the binding affinities of all four CROSS 
      proteins to a panel of two wild-type and six chimeric chemokines. These 
      complementation studies indicate the regions of the chemokines that interact with 
      each element of the receptors, allowing us to deduce the orientations of the 
      receptor extracellular elements relative to the bound chemokines.
FAU - Datta-Mannan, Amita
AU  - Datta-Mannan A
AD  - Department of Chemistry, Indiana University, Bloomington, Indiana 47405-0001, 
      USA.
FAU - Stone, Martin J
AU  - Stone MJ
LA  - eng
GR  - GM55055/GM/NIGMS NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Chemokine CCL11)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Chemokines, CC)
RN  - 0 (Receptors, CCR2)
RN  - 0 (Receptors, CCR3)
RN  - 0 (Receptors, Chemokine)
RN  - 0 (Recombinant Fusion Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Chemokine CCL11
MH  - Chemokine CCL2/metabolism
MH  - Chemokines, CC/genetics/*metabolism
MH  - Models, Molecular
MH  - *Molecular Mimicry
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Protein Binding
MH  - Receptors, CCR2
MH  - Receptors, CCR3
MH  - Receptors, Chemokine/genetics/*metabolism
MH  - Recombinant Fusion Proteins/genetics/*metabolism
MH  - Solubility
EDAT- 2004/11/17 09:00
MHDA- 2005/01/19 09:00
CRDT- 2004/11/17 09:00
PHST- 2004/11/17 09:00 [pubmed]
PHST- 2005/01/19 09:00 [medline]
PHST- 2004/11/17 09:00 [entrez]
AID - 10.1021/bi048990e [doi]
PST - ppublish
SO  - Biochemistry. 2004 Nov 23;43(46):14602-11. doi: 10.1021/bi048990e.