Signal Transduction and Ligand Specificity of the Human Monocyte Chemoattractant Protein-1 Receptor in Transfected Embryonic Kidney Cells (∗)

We have examined the ligand specificity and signal transduction pathways of a recently cloned receptor for monocyte chemoattractant protein-1 (MCP-1). In human 293 cells stably transfected with the MCP-1 receptor, MCP-1 bound specifically with high affinity (Kd = 260 pM) and induced a rapid mobiliza...

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Bibliographic Details
Published in:The Journal of biological chemistry 1995-03, Vol.270 (11), p.5786-5792
Main Authors: Myers, Scott J., Wong, Lu Min, Charo, Israel F.
Format: Article
Language:English
Subjects:
Adenylyl Cyclase Inhibitors
Binding, Competitive
Calcium - metabolism
Cell Line
Chemokine CCL2
Chemotactic Factors - metabolism
Cytokines - pharmacology
Embryo, Mammalian
Humans
Iodine Radioisotopes
Kidney
Kinetics
Ligands
Lymphokines - pharmacology
Receptors, CCR2
Receptors, Chemokine
Receptors, Cytokine - metabolism
Recombinant Proteins - metabolism
Signal Transduction
Substrate Specificity
Transfection
Type C Phospholipases - metabolism
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Summary:We have examined the ligand specificity and signal transduction pathways of a recently cloned receptor for monocyte chemoattractant protein-1 (MCP-1). In human 293 cells stably transfected with the MCP-1 receptor, MCP-1 bound specifically with high affinity (Kd = 260 pM) and induced a rapid mobilization of calcium from intracellular stores. The closely related chemokines MIP-1α, MIP-1β, RANTES, interleukin 8 (IL-8), and Gro-α were inactive at concentrations as high as 300 nM. Activation of the MCP-1 receptor potently inhibited adenylyl cyclase with an IC50 = 90 pM. Activation of the MIP-1α/RANTES receptor also mediated inhibition of adenylyl cyclase activity but with a different pharmacological profile: MIP-1α (110 pM, IC50), RANTES (140 pM), MIP-1β (10 nM), and MCP-1 (820 nM). Mobilization of intracellular calcium and inhibition of adenylyl cyclase were blocked by pertussis toxin, suggesting that the MCP-1 receptor coupled to Gαi. These results demonstrate that the MCP-1 receptor binds and signals in response to picomolar concentrations of MCP-1 in a highly specific manner. Signaling was manifested as mobilization of intracellular calcium and inhibition of adenylyl cyclase and was mediated by a pertussis toxin-sensitive G-protein(s).
ISSN:0021-9258
1083-351X
DOI:10.1074/jbc.270.11.5786