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 Ca2+-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 12nM 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.
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September 2001
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Research Article|
September 10 2001
Multimerization of monocyte chemoattractant protein-1 is not required for glycosaminoglycan-dependent transendothelial chemotaxis
Simi ALI;
Simi ALI
Immunobiology Group, Department of Surgery, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, U.K.
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Adrian C. V. PALMER;
Adrian C. V. PALMER
Immunobiology Group, Department of Surgery, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, U.K.
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Sarah J. FRITCHLEY;
Sarah J. FRITCHLEY
Immunobiology Group, Department of Surgery, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, U.K.
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Yvonne MALEY;
Yvonne MALEY
Immunobiology Group, Department of Surgery, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, U.K.
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John A. KIRBY
John A. KIRBY
1
Immunobiology Group, Department of Surgery, The Medical School, University of Newcastle, Newcastle upon Tyne NE2 4HH, U.K.
1To whom correspondence should be addressed (e-mail [email protected]).
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Publisher: Portland Press Ltd
Received:
January 23 2001
Revision Received:
June 06 2001
Accepted:
July 10 2001
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2001
2001
Biochem J (2001) 358 (3): 737–745.
Article history
Received:
January 23 2001
Revision Received:
June 06 2001
Accepted:
July 10 2001
Citation
Simi ALI, Adrian C. V. PALMER, Sarah J. FRITCHLEY, Yvonne MALEY, John A. KIRBY; Multimerization of monocyte chemoattractant protein-1 is not required for glycosaminoglycan-dependent transendothelial chemotaxis. Biochem J 15 September 2001; 358 (3): 737–745. doi: https://doi.org/10.1042/bj3580737
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