Characterization of a chimeric chemokine as a specific ligand for ACKR3
Characterization of a chimeric chemokine as a specific ligand for ACKR3
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DOI：
10.1002/jlb.2ma1217-509r
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发表时间：
2018-08-01
影响因子：
5.5
通讯作者：
Thelen, Marcus
中科院分区：
文献类型：
--
作者：
Ameti, Rafet;Melgrati, Serena;Thelen, Marcus
趋化因子是一种小的趋化细胞因子，通过与其同源趋化因子受体结合来协调细胞迁移。虽然趋化因子介导的典型G蛋白偶联趋化因子受体的刺激导致细胞迁移，但趋化因子与非典型趋化因子受体（ACKR）的结合不诱导典型信号传导。ACKR被认为是重要的趋化因子清除剂，可以产生梯度，帮助引导细胞到炎症部位或其免疫小生境。合成趋化因子在过去已被用于研究和解码趋化因子-受体相互作用。表征特定的趋化因子-ACKR相互作用是具有挑战性的，因为趋化因子结合多种受体;例如，ACKR 3配体CXCL 12和CXCL 11分别结合经典受体CXCR 4和CXCR 3。在这里，我们提出了趋化因子-N样嵌合体的工程设计，它选择性地结合ACKR 3。在C末端添加ybbR 13标签允许用过多的荧光染料进行位点特异性酶标记。嵌合体由CXCL 11的N-末端和CXCL 12的主体和C-末端组成，并且以高亲和力选择性地与ACKR 3相互作用，同时不干扰CXCL 11和CXCL 12与其同源受体的结合。我们进一步提供了证据表明，嵌合体可用于研究体内ACKR 3功能。
Chemokines, small chemotactic cytokines, orchestrate cell migration by binding to their cognate chemokine receptors. While chemokine-mediated stimulation of typical G-protein-coupled chemokine receptors leads to cell migration, binding of chemokines to atypical chemokine receptors (ACKRs) does not induce canonical signaling. ACKRs are considered important chemokine scavengers, that can create gradients which help direct cells to sites of inflammation or to their immunological niches. Synthetic chemokines have been used in the past to study and decode chemokine-receptor interactions. Characterizing specific chemokine-ACKRs interactions is challenging because the chemokines bindmultiple receptors; for example, the ACKR3 ligands CXCL12 and CXCL11 bind to the canonical receptors CXCR4 and CXCR3, respectively. Here, we present the engineering of a chemokine-N like chimera, which selectively binds to ACKR3. The addition of a ybbR13 tag at the C-terminus allows site specific enzymatic labeling with a plethora of fluorescent dyes. The chimera is composed of the N-terminus of CXCL11 and the main body and C-terminus of CXCL12 and selectively interacts with ACKR3 with high affinity, while not interfering with binding of CXCL11 and CXCL12 to their cognate receptors. We further provide evidence that the chimera can be used to study ACKR3 function in vivo.