Functional Characterization of Three Mouse Formyl Peptide Receptors
Functional Characterization of Three Mouse Formyl Peptide Receptors
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DOI：
10.1124/mol.112.081315
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发表时间：
2013-02-01
影响因子：
3.6
通讯作者：
Ye, Richard D.
中科院分区：
文献类型：
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作者：
He, Hui-Qiong;Liao, Dan;Ye, Richard D.
人类甲酰肽受体（FPRs）和它们的小鼠对应物之间的进化关系和功能相关性仍然不完全清楚。我们研究了小鼠甲酰肽受体亚家族（mFXR）的三个成员，发现它们在激动剂偏好和细胞分布方面存在差异。当在转染的大鼠嗜碱性白血病（RBL-2 H3）细胞中稳定表达时，mFpr 1很容易被单核细胞增生李斯特菌（fMIVTLF）、金黄色葡萄球菌（fMIFL）和线粒体（fMMYALF）的N-甲酰化肽激活。相比之下，大肠杆菌衍生的fMLF的效力低1000倍。上述肽对mFpr 2的有效性低得多，mFpr 2对合成的六肽WKYMVm、合成的激动剂Quin-C1（取代的喹唑啉酮）和化合物43（亚硝基化的吡唑啉酮衍生物）的反应更好。饱和结合试验表明，mFpr 1和mFpr 2在细胞表面上以相似的水平表达，尽管它们对N-甲酰基-Met-Leu-Phe-Ile-Ile-Lys-异硫氰酸荧光素的亲和力变化超过1000倍[mFpr 1的解离常数（Kd）值为2.8 nM，mFpr 2的解离常数（Kd）值为4.8 μ M]。与这些受体相反，mFpr-rs 1对所有先前提到的测试肽的反应都很差。荧光显微镜观察显示mFpr-rs 1在细胞内的分布模式。基于这些结果，我们得出结论，mFpr 1是人FPR 1的直系同源物，具有人FPR 2/ALX的某些药理学特性，而mFpr 2对甲酰基肽的亲和力低得多。mFpr-rs 1的细胞内分布表明与人FPR 3的进化相关性。
The evolutionary relationship and functional correlation between human formyl peptide receptors (FPRs) and their mouse counterparts remain incompletely understood. We examined three members of the mouse formyl peptide receptor subfamily (mFprs) and found that they differ in agonist preference and cellular distributions. When stably expressed in transfected rat basophilic leukemia (RBL-2H3) cells, mFpr1 was readily activated by N-formylated peptides derived from Listeria monocytogenes (fMIVTLF), Staphylococcus aureus (fMIFL), and mitochondria (fMMYALF). In contrast, the Escherichia coli-derived fMLF was 1000-fold less potent. The aforementioned peptides were much less efficacious at mFpr2, which responded better to the synthetic hexapeptide WKYMVm, the synthetic agonists Quin-C1 (a substituted quinazolinone), and compound 43 (a nitrosylated pyrazolone derivative). Saturation binding assays showed that mFpr1 and mFpr2 were expressed at similar levels on the cell surface, although their affinity for N-formyl-Met-Leu-Phe-Ile-Ile-Lys-fluorescein isothiocyanate varied by more than 1000-fold [dissociation constant (K-d) values of 2.8 nM for mFpr1 and 4.8 mu M for mFpr2]). Contrary to these receptors, mFpr-rs1 responded poorly to all the previously mentioned peptides that were tested. Fluorescent microscopy revealed an intracellular distribution pattern of mFpr-rs1. On the basis of these results, we conclude that mFpr1 is an ortholog of human FPR1 with certain pharmacologic properties of human FPR2/ALX, whereas mFpr2 has much lower affinity for formyl peptides. The intracellular distribution of mFpr-rs1 suggests an evolutionary correlation with human FPR3.