PMID- 22396538
OWN - NLM
STAT- MEDLINE
DCOM- 20120702
LR  - 20211021
IS  - 1083-351X (Electronic)
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 287
IP  - 18
DP  - 2012 Apr 27
TI  - Design and receptor interactions of obligate dimeric mutant of chemokine monocyte 
      chemoattractant protein-1 (MCP-1).
PG  - 14692-702
LID - 10.1074/jbc.M111.334201 [doi]
AB  - Chemokine-receptor interactions regulate leukocyte trafficking during 
      inflammation. CC chemokines exist in equilibrium between monomeric and dimeric 
      forms. Although the monomers can activate chemokine receptors, dimerization is 
      required for leukocyte recruitment in vivo, and it remains controversial whether 
      dimeric CC chemokines can bind and activate their receptors. We have developed an 
      obligate dimeric mutant of the chemokine monocyte chemoattractant protein-1 
      (MCP-1) by substituting Thr(10) at the dimer interface with Cys. Biophysical 
      analysis showed that MCP-1(T10C) forms a covalent dimer with similar structure to 
      the wild type MCP-1 dimer. Initial cell-based assays indicated that MCP-1(T10C) 
      could activate chemokine receptor CCR2 with potency reduced 1 to 2 orders of 
      magnitude relative to wild type MCP-1. However, analysis of size exclusion 
      chromatography fractions demonstrated that the observed activity was due to a 
      small proportion of MCP-1(T10C) being monomeric and highly potent, whereas the 
      majority dimeric form could neither bind nor activate CCR2 at concentrations up 
      to 1 muM. These observations help to reconcile previous conflicting results and 
      indicate that dimeric CC chemokines do not bind to their receptors with 
      affinities approaching those of the corresponding monomeric chemokines.
FAU - Tan, Joshua H Y
AU  - Tan JH
AD  - Department of Biochemistry and Molecular Biology, Monash University, Clayton, 
      Victoria 3800, Australia.
FAU - Canals, Meritxell
AU  - Canals M
FAU - Ludeman, Justin P
AU  - Ludeman JP
FAU - Wedderburn, Jamie
AU  - Wedderburn J
FAU - Boston, Christopher
AU  - Boston C
FAU - Butler, Stephen J
AU  - Butler SJ
FAU - Carrick, Ann Marie
AU  - Carrick AM
FAU - Parody, Todd R
AU  - Parody TR
FAU - Taleski, Deni
AU  - Taleski D
FAU - Christopoulos, Arthur
AU  - Christopoulos A
FAU - Payne, Richard J
AU  - Payne RJ
FAU - Stone, Martin J
AU  - Stone MJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120306
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (CCL2 protein, human)
RN  - 0 (CCR2 protein, human)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Receptors, CCR2)
SB  - IM
MH  - Amino Acid Substitution
MH  - Cell Line
MH  - Chemokine CCL2/chemistry/genetics/*metabolism
MH  - Humans
MH  - *Mutation, Missense
MH  - *Protein Multimerization
MH  - Receptors, CCR2/chemistry/genetics/metabolism
PMC - PMC3340267
EDAT- 2012/03/08 06:00
MHDA- 2012/07/03 06:00
PMCR- 2013/04/27
CRDT- 2012/03/08 06:00
PHST- 2012/03/08 06:00 [entrez]
PHST- 2012/03/08 06:00 [pubmed]
PHST- 2012/07/03 06:00 [medline]
PHST- 2013/04/27 00:00 [pmc-release]
AID - S0021-9258(20)47961-X [pii]
AID - M111.334201 [pii]
AID - 10.1074/jbc.M111.334201 [doi]
PST - ppublish
SO  - J Biol Chem. 2012 Apr 27;287(18):14692-702. doi: 10.1074/jbc.M111.334201. Epub 
      2012 Mar 6.