FEBS Letters | |
Characterisation of the RANTES/MIP‐1α receptor (CC CKR‐1) stably transfected in HEK 293 cells and the recombinant ligands | |
Montjovent, Marc-Olivier1  Hoogewerf, Arlene1  Power, Christine A.1  Wells, Timothy N.C.1  Borlat, Frederic1  Proudfoot, Amanda E.I.1  | |
[1] Glaxo Institute for Molecular Biology, 14, Ch. des Aulx, 1228 Plan-les-Ouates, Geneva, Switzerland | |
关键词: Recombinant chemokine; Chemotaxis; RANTES/MIP-1α receptor; HEK 293 transfectant; RANTES; Regulated on Activation; Normal T expressed and Secreted; MIP-1α; macrophage inflammatory protein-1α; MCP-1; monocyte chemotactic protein-1; Il-8; interleukin-8; IPTG; isopropylthiogalactoside; EC50; concentration of agonist giving half the maximal response; IC50; concentration of inhibitor giving half the maximal inhibition; | |
DOI : 10.1016/0014-5793(95)01235-X | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
The CC chemokines RANTES and MIP-1α are known to activate certain leucocytes and leucocytic cell lines. We have produced and fully characterised the recombinant proteins expressed in E. coli. They induce chemotaxis of the pro-monocytic cell line, THP-1 and T cells. THP-1 cells express three of the known CC chemokine receptors. In order to study the activation of a single receptor, we have expressed the shared receptor (CC CKR-1) for RANTES and MIP-1α stably in the HEK 293 cell line. We have examined the effects of RANTES and MIP-1α on the CC CKR-1 transfectants by equilibrium binding studies and in a chemotaxis assay. RANTES competes for [125I]RANTES with an IC50 of 0.6 ± 0.23 nM, whereas MIP-1α competes for its radiolabelled counterpart with an IC50 of 10 ± 1.6 nM in the transfectants. These affinities are the same as those measured on the THP-1 cell line. The stably transfected HEK 293 cells respond to both these chemokines in the chemotaxis assay with the same EC50 values as those measured for THP-1 cells. This indicates that this cellular response can be mediated through the CC CKR-1 receptor.
【 授权许可】
Unknown
【 预 览 】
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