The synthetic peptide Trp-Lys-Tyr-Met-Val-D-Met is a potent chemotactic agonist for mouse formyl peptide receptor
The synthetic peptide Trp-Lys-Tyr-Met-Val-D-Met is a potent chemotactic agonist for mouse formyl peptide receptor
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DOI：
10.4049/jimmunol.165.8.4598
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发表时间：
2000-10-15
影响因子：
4.4
通讯作者：
Ye, RD
中科院分区：
文献类型：
--
作者：
He, R;Tan, L;Ye, RD
甲酰基肽是有效的中性粒细胞化学引诱剂。在人和家兔中，甲酰基肽受体（FPR）与n -甲酰基met - leu - phe （fMLF）具有高亲和力（K-d约为1 nM），小鼠FPR （mFPR）是fMLF的低亲和力受体（K-d约为100 nM）；因此，可能存在该受体的其他激动剂。利用转染mFPR的大鼠嗜碱性白血病细胞，我们发现最近发现的合成肽Trp-Lys-Tyr-Met-Val-D-Met （WKYMVm）是一种有效的mFPR激动剂，WKYMVm诱导钙动员的EC50为1.2-1.5 nM， I nM的WKYMVm达到最佳趋化性，但需要100 nM的fMLF。WKYMVm在50 nM下刺激了丝裂原活化蛋白激酶细胞外信号相关激酶1和2的快速有效磷酸化，百日毒仅部分阻断了受刺激的mFPR细胞中钙的动员和肌醇1,4,5 -三磷酸的产生，这表明该受体可能与除Gi和Go以外的G α蛋白偶联。竞争性结合和脱敏数据表明，这两种肽与同一受体相互作用，但可能使用不重叠的结合位点，因为WKYMVm不能有效地取代与mFPR结合的[H-3]fMLF。这些结果为mFPR存在另一种有效的激动剂提供了证据，并表明WKYMVm可能用于探测与小鼠甲酰基肽受体同源物的配体受体相互作用。
Formyl peptides are potent neutrophil chemoattractants. In humans and rabbits, the formyl peptide receptor (FPR) binds N-formyl-Met-Leu-Phe (fMLF) with high affinity (K-d approximate to 1 nM), The mouse FPR (mFPR) is a low-affinity receptor for fMLF (K-d approximate to 100 nM); therefore, other agonists for this receptor may exist. Using mFPR-transfected rat basophilic leukemia cells, we found that a recently identified synthetic peptide Trp-Lys-Tyr-Met-Val-D-Met (WKYMVm) is a potent agonist for mFPR, WKYMVm induced calcium mobilization with an EC50 of 1.2-1.5 nM, Optimal chemotaxis was achieved with I nM of WKYMVm, but it required 100 nM of fMLF. WKYMVm stimulated rapid and potent phosphorylation of the mitogen-activated protein kinases extracellular signal-related kinases 1 and 2 when used at 50 nM, Pertussis toxin only partially blocked calcium mobilization and production of inositol I,4,5-trisphosphate in the stimulated mFPR cells, suggesting the possibility that this receptor couples to G alpha proteins other than Gi and Go. Competitive binding and desensitization data suggest that both peptides interact with the same receptor but may use nonoverlapping binding sites because WKYMVm was unable to effectively displace [H-3]fMLF bound to mFPR, These results provide evidence for the presence of an alternative potent agonist for mFPR, and suggest a potential usage of WKYMVm for probing the ligand-receptor interactions with the murine formyl peptide receptor homologs.