Analysis of the linear epitope for Fc-binding on the mouse IgG Fc receptor (moFcγRI) by synthetic peptide.
Analysis of the linear epitope for Fc-binding on the mouse IgG Fc receptor (moFcγRI) by synthetic peptide.
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DOI：
10.4238/2014.june.18.7
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发表时间：
2014-06
期刊：
影响因子：
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通讯作者：
Wang Fy;Guo Jq;Gaiping Zhang
中科院分区：
文献类型：
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作者：
Wang Fy;Guo Jq;Gaiping Zhang
为了确定Fc与小鼠免疫球蛋白G(Ig G)Fc受体(mOFCγRI)结合的线性表位，从mOFCγRI的膜-远端胞外域(EC2)出发，合成了与人FcγRI(huFcγRI)和huFcγRII同源区域相对应的多肽。用斑点印迹法检测了6个多肽。结果表明，MoRI3多肽(CVFYRNGKSFQFS)能有效地与小鼠免疫球蛋白结合。竞争性酶联免疫吸附试验表明，moRI3多肽的IC50值为38.03 mM。MoRI3多肽能显著抑制小鼠免疫球蛋白与COS-7细胞的结合，IC50值为72.68 mM。根据huFcγRI和huFcγRII的预测，mOFCγRI的免疫球蛋白结合区也定位于EC2结构域的C‘-E环。我们预测γRI的最小有效免疫球蛋白结合区可能是153SFQFSS158。还描述了免疫球蛋白与小鼠FcγR结合的线性表位。因此，我们产生了一种以该受体类识别配体的基本方面为靶点的多肽。
To identify the linear epitope for Fc-binding to the mouse immunoglobulin G (IgG) Fc receptor (moFcγRI), peptides derived from the membrane-distal extracellular domain (EC2) of moFcγRI, corresponding to the homologous region of human FcγRI (huFcγRI) and huFcγRII, were synthesized. Using a dot-blot assay, six peptides were tested. The results showed that the moRI3 peptide (CVFYRNGKSFQFS) could combine with mouse IgG efficiently. A competitive enzyme-linked immunosorbent assay (ELISA) showed that the IC50 value of the moRI3 peptide was 38.03 mM. The moRI3 peptide could inhibit the combination of mouse IgG to the transfected COS 7 cells significantly with an IC50 value of 72.68 mM. The IgG-binding region of moFcγRI was also localized in the C'-E loop of the EC2 domain as predicted according to huFcγRI and huFcγRII. We predicted that the minimum effective IgG-binding region of moFcγRI may be the peptide 153SFQFSS158. The linear epitope for immunoglobulin-binding to mouse FcγR is also described. Thus, we generated a peptide that targets a fundamental aspect of ligand recognition by this receptor class.