PMID- 11535134
OWN - NLM
STAT- MEDLINE
DCOM- 20011025
LR  - 20190501
IS  - 0264-6021 (Print)
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 358
IP  - Pt 3
DP  - 2001 Sep 15
TI  - Multimerization of monocyte chemoattractant protein-1 is not required for 
      glycosaminoglycan-dependent transendothelial chemotaxis.
PG  - 737-45
AB  - Chemokines interact with specific G-protein-coupled cell-surface receptors and 
      with glycosaminoglycans (GAGs), such as heparan sulphate. Although chemokines 
      often form multimers in solution, this process may be enhanced following 
      interaction with GAGs on the cell surface, or within the extracellular matrix. 
      However, the significance of multimerization for chemokine function remains 
      controversial. In the present study, a fusion protein was prepared between the 
      prototypical human CC chemokine, monocyte chemoattractant protein-1 (MCP-1; also 
      known as CCL-2) and a large secreted placental alkaline phosphatase (SEAP) 
      moiety. This fusion protein (MCP-1-SEAP) remained monomeric under conditions that 
      promote oligomerization of the native chemokine. Radioligand binding showed that 
      both native MCP-1 and MCP-1-SEAP competed for the same site on the surface of 
      HEK-293 cells expressing the CCR2b chemokine receptor. The interaction between 
      either chemokine species and endothelial cell surface GAGs was antagonized by the 
      addition of the heparan sulphate-like molecule, heparin. Both MCP-1 and 
      MCP-1-SEAP induced a Ca(2+)-flux in the THP-1 monocytic cell line, and were 
      equally effective at promoting transendothelial chemotaxis of mononuclear immune 
      cells, with maximal migration being produced by treatment with 12 nM of either 
      species. In each case this chemotactic response was almost completely antagonized 
      by the addition of heparin. The importance of interaction between either native 
      MCP-1 or MCP-1-SEAP and cell-surface GAGs for transcellular migration was 
      demonstrated by the almost complete absence of leucocyte chemotaxis across 
      monolayers of GAG-deficient mutant cells. In summary, this study shows that 
      multimerization is neither necessary for, nor potentiates, the biological 
      activity of MCP-1. However, the results do clearly demonstrate the importance of 
      the interaction between MCP-1 and cell-surface heparan sulphate for 
      transmonolayer leucocyte chemotaxis.
FAU - Ali, S
AU  - Ali S
AD  - Immunobiology Group, Department of Surgery, The Medical School, University of 
      Newcastle, Newcastle upon Tyne NE2 4HH, UK.
FAU - Palmer, A C
AU  - Palmer AC
FAU - Fritchley, S J
AU  - Fritchley SJ
FAU - Maley, Y
AU  - Maley Y
FAU - Kirby, J A
AU  - Kirby JA
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (CCR2 protein, human)
RN  - 0 (Ccr2 protein, mouse)
RN  - 0 (Chemokine CCL2)
RN  - 0 (Glycosaminoglycans)
RN  - 0 (Ligands)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Receptors, CCR2)
RN  - 0 (Receptors, Chemokine)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 59880-97-6 (N-Formylmethionine Leucyl-Phenylalanine)
RN  - 9005-49-6 (Heparin)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
RN  - SY7Q814VUP (Calcium)
SB  - IM
MH  - 3T3 Cells
MH  - Alkaline Phosphatase/genetics/metabolism
MH  - Animals
MH  - Binding Sites
MH  - CHO Cells
MH  - Calcium/metabolism
MH  - Cell Line
MH  - Cell Membrane/physiology
MH  - Cells, Cultured
MH  - Chemokine CCL2/genetics/pharmacology/*physiology
MH  - *Chemotaxis, Leukocyte/drug effects
MH  - Cloning, Molecular
MH  - Cricetinae
MH  - Endothelium, Vascular/cytology/*physiology
MH  - Female
MH  - Glycosaminoglycans/*pharmacology
MH  - Heparin/pharmacology
MH  - Humans
MH  - Ligands
MH  - Macromolecular Substances
MH  - Mice
MH  - Monocytes/*physiology
MH  - N-Formylmethionine Leucyl-Phenylalanine/pharmacology
MH  - Placenta/enzymology
MH  - Pregnancy
MH  - Receptors, CCR2
MH  - Receptors, Chemokine/genetics/*physiology
MH  - Recombinant Fusion Proteins/metabolism
MH  - Recombinant Proteins/metabolism
MH  - Transfection
PMC - PMC1222107
EDAT- 2001/09/06 10:00
MHDA- 2001/10/26 10:01
PMCR- 2002/03/15
CRDT- 2001/09/06 10:00
PHST- 2001/09/06 10:00 [pubmed]
PHST- 2001/10/26 10:01 [medline]
PHST- 2001/09/06 10:00 [entrez]
PHST- 2002/03/15 00:00 [pmc-release]
AID - 10.1042/0264-6021:3580737 [doi]
PST - ppublish
SO  - Biochem J. 2001 Sep 15;358(Pt 3):737-45. doi: 10.1042/0264-6021:3580737.