Two C-terminal peptides of human CKLF1 interact with the chemokine receptor CCR4
Two C-terminal peptides of human CKLF1 interact with the chemokine receptor CCR4
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人 CKLF1 的两个 C 端肽与趋化因子受体 CCR4 相互作用
DOI：
10.1016/j.biocel.2007.10.028
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发表时间：
2008-01-01
影响因子：
4
通讯作者：
Ma, Dalong
中科院分区：
文献类型：
--
作者：
Wang, Ying;Zhang, Yingmei;Ma, Dalong
人趋化因子样因子1 （CKLF1）对白细胞具有趋化作用。先前的研究表明，CKLF1是人CC趋化因子受体4 （CCR4）的功能性配体。本研究对分泌的CKLF1蛋白进行n端氨基酸测序，发现其至少含有C27和C19两条肽段。为了研究C27或C19是否通过CCR4发挥作用，我们在CCR4转染的HEK293细胞或hu78细胞中化学合成C27和C19，并通过趋化性、钙动员和受体内化实验分析C27和C19。趋化实验表明，C27对转染ccr4的HEK293细胞和hu78细胞具有趋化作用，而C19对hu78细胞的趋化作用较弱。C27-或c19诱导的趋化性被百日咳毒素消除，提示Gi/o通路的参与。C27-或c19诱导的趋化作用也可被CCR4拮抗剂抑制，该拮抗剂对CCR4具有良好的结合效力、优异的趋化抑制活性和选择性。它们的趋化活性特别涉及CCR4。经TARC/CCL17预孵育后，ccr4转染的HEK293细胞对C27或C19的趋化反应被明显抑制。TARC/CCL17能有效脱敏C27或C19诱导的钙动员。同样，C27或C19也使ccr4转染的HEK293细胞对TARC/CCL17的钙动员和趋化脱敏，这表明它们可能与一个共同的受体相互作用。C27-和c19均诱导CCR4-EGFP明显内化。这些结果证实了CKLF1、C27和C19的分泌肽可通过CCR4进行功能激活。(C) 2007 Elsevier Ltd.版权所有。
Human chemokine-like factor 1 (CKLF1) exhibits chemotactic effects on leukocytes. A previous study demonstrated that CKLF1 is a functional ligand for human CC chemokine receptor 4 (CCR4). In this study, N-terminal amino acid sequencing of secreted CKLF1 protein showed that it contains at least two peptides, C27 and C19. To examine whether C27 or C19 play a role via CCR4, C27 and C 19 were chemically synthesized and analyzed by chemotaxis, calcium mobilization, and receptor internalization assays in CCR4-tranfected HEK293 cells or Hut78 cells. The chemotaxis assay showed that C27 could induce chemotaxis to CCR4-transfected HEK293 cells or Hut78 cells while C19 had weaker chemotactic activity, especially in Hut78 cells. C27- or C19-induced chemotaxis was abolished by pertussis toxin, suggesting the involvement of a Gi/o pathway. C27- or C19-induced chemotaxis was also inhibited by an antagonist of CCR4 that show good binding potency, excellent chemotaxis inhibitory activity and selectivity toward CCR4, suggesting. that their chemotactic activity specifically involved CCR4. The chemotactic response of CCR4-tranfected HEK293 cells to C27 or C19 was markedly inhibited by preincubation with TARC/CCL17. TARC/CCL17 effectively desensitized the calcium mobilization induced by C27 or C19. Similarly, both of C27 or C19 also desensitized the calcium mobilization and chemotaxis of CCR4-tranfected HEK293 cells in response to TARC/CCL17, suggesting that they might interact with a common receptor. Both C27- and C19-induced clear internalization of CCR4-EGFP. These results confirm that the secreted peptides of CKLF1, C27 and C19, have functional activation via CCR4. (C) 2007 Elsevier Ltd. All rights reserved.