Analysis of CS5 co-receptor molecule that specifically inhibits chemotaxis of polymorphonuclear leukocytes

特异性抑制多形核白细胞趋化性的CS5共受体分子分析

基本信息

批准号：
12470058
负责人：
YAMAMOTO Tetsuro
金额：
$ 7.62万
依托单位：
Kumamoto University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470058/
关键词：
ribosomal protein leukocyte chemotaxis C5a receptor apoptosis transglutaminase inflammation antagonist C5a 光アフィニティ標識

项目摘要

The cross-linked dimer of RP S19 attracted monocytes as an agonist of C5a receptor. On the other hand, the same molecule inhibited the chemotaxis of polymorphonuclear leukocytes (PMN) as an antagonist of the C5a receptor. We initially identified the ligand moieties of the RP S19 dimer to the C5a receptor of monocytes. The RP S19 dimer bound to the C5a receptor by a two-step mechanism. The first ligand and second ligand moieties were identified to be-Lys41-His42-Lys43- and -Leu131-Asp132-Arg133-, respectively.Next, we identified a switch moiety on the RPS19 dimer molecule, which converted the agonist function to the antagonist in the C5a receptor-mediated chemotaxis of PMN. The switch moiety was identified to be from Ile134 to Lys144 (IAGQVAAANKK), which was present following to the second ligand moiety.We made a hypothesis that PMN had a molecule near the C5a receptor, which negatively regulated the intracellular signaling raised by the activated C5a receptor. Then we have been trying to identify the co-receptor molecule. We have prepared a subtraction cDNA library between a PMN cDNA pool and a monocyte cDNA pool. We have tried to prepare an analogue peptide with the second ligand moiety and the switch moiety that bore a fluorescence-labeled photosensitive chemical modifier. However, we have not succeeded to identify the co-receptor molecule of the PMN C5a receptor.

RP S19的交联二聚体吸引了单核细胞作为C5A受体的激动剂。另一方面，相同的分子抑制了多形核白细胞（PMN）作为C5A受体的拮抗剂的趋化性。我们最初将RP S19二聚体的配体部分鉴定为单核细胞的C5A受体。 RP S19二聚体通过两步机制结合到C5A受体。 The first ligand and second ligand moieties were identified to be-Lys41-His42-Lys43- and -Leu131-Asp132-Arg133-, respectively.Next, we identified a switch moiety on the RPS19 dimer molecule, which converted the agonist function to the antagonist in the C5a receptor-mediated chemotaxis of PMN.开关部分被确定为从ILE134到LYS144（IAGQVAAANKK），随后存在于第二个配体部分。我们做出了一个假设，即PMN在C5A受体附近具有分子，该分子在激活的C5A受体中对细胞内信号进行了负调控。然后，我们一直在尝试识别共受体分子。我们已经在PMN cDNA池和单核细胞cDNA池之间制备了减法cDNA文库。我们试图用第二个配体部分和带有荧光标记的光敏化学修饰剂的开关部分制备模拟肽。但是，我们尚未成功识别PMN C5A受体的共受体分子。