The formylpeptide chemotactic receptor on rabbit peritoneal neutrophils. I. Evidence for two binding sites with different affinities.
The formylpeptide chemotactic receptor on rabbit peritoneal neutrophils. I. Evidence for two binding sites with different affinities.
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兔腹膜中性粒细胞上的甲酰肽趋化受体。
DOI：
10.4049/jimmunol.129.4.1608
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发表时间：
1982
影响因子：
4.4
通讯作者：
E. Becker
中科院分区：
文献类型：
--
作者：
W. Mackin;C. Huang;E. Becker
在4 ℃硅油离心试验中测量F Met-Leu-[3 H]Phe和f Nle-Leu-[3 H]Phe与兔腹膜中性粒细胞和纯化膜的结合，并通过LIGAND计算机程序分析结果，该程序允许分析配体与多种结合位点的结合。肽结合完整的中性粒细胞的配体分析表明，f Met-Leu-[3 H]Phe和f Nle-Leu-[3 H]Phe检测到两个群体的结合位点。f Met-Leu-[3 H]Phe结合的表观Kd值分别为1.6 +/- 1.0 X 10（-9）M和2.2 +/- 0.9 X 10（-8）M，f Nle-Leu-[3 H]Phe结合的表观Kd值分别为3.1 +/- 0.2 X 10（-9）M和1.2 +/- 0.6 X 10（-7）M。此外，在全细胞上检测到的较高亲和力位点占总位点的约15 - 30%。两个群体的结合位点也检测到纯化的中性粒细胞质膜的放射性标记的趋化肽。肽与纯化膜结合的配体分析得出，f Met-Leu-[3 H] Phe结合的表观Kd值为5.0 +/- 2.5 X 10（-10）M和4.8 +/- 0.6 X 10（-8）M，f Nle-Leu-[3 H] Phe为4.7 +/- 4.2 X 10（-10）M和3.0 +/- 1.3 X 10（-8）M。在纯化膜上由f Met-Leu-[3 H]Phe和f Nle-Leu-[3 H]Phe检测到的高亲和力位点的百分比为检测到的总位点的1%至5%。这些数据是一致的，要么与存在两个独立的结合位点的甲酰基肽对兔中性粒细胞或受体负协同性。
F Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe binding to rabbit peritoneal neutrophils and purified membranes were measured at 4 degrees C silicone oil centrifugation assays, and the results were analyzed by the LIGAND computer program, which permits analysis of ligand binding to multiple classes of binding sites. LIGAND analysis of peptide binding to intact neutrophil indicated that both f Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe detected two population of binding sites. The apparent Kd values for f Met-Leu-[3H]Phe binding were 1.6 +/- 1.0 X 10(-9) M and 2.2 +/- 0.9 X 10(-8) M, respectively, and 3.1 +/- 0.2 X 10(-9) M and 1.2 +/- 0.6 X 10(-7) M for f Nle-Leu-[3H]Phe. Furthermore, the higher affinity sites detected on whole cells comprised approximately 15 to 30% of the total sites. Two populations of binding sites were also detected on purified neutrophil plasma membranes by both radiolabeled chemotactic peptides. LIGAND analysis of peptide binding to purified membranes yielded apparent Kd values of 5.0 +/- 2.5 X 10(-10) M and 4.8 +/- 0.6 X 10(-8) M for f Met-Leu-[3H]Phe binding, and 4.7 +/- 4.2 X 10(-10) M and 3.0 +/- 1.3 X 10(-8) M for f Nle-Leu-[3H]Phe. The percentage of higher affinity sites detected by f Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe on purified membranes were 1 to 5% of the total sites detected. These data are consistent either with the existence of two independent binding sites for formylpeptides on rabbit neutrophils or receptor negative cooperativity.