Novel Designs of Single-chain MHC I/peptide Complex for the Magnetosome Display System
Novel Designs of Single-chain MHC I/peptide Complex for the Magnetosome Display System
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用于磁小体显示系统的单链 MHC I/肽复合物的新颖设计
DOI：
10.1093/protein/gzu056
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Tomoko Yoshino
中科院分区：
文献类型：
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作者：
Toru Honda;Yoshiaki Maeda;Takayuki Yasuda;Tsuyoshi Tanaka;Tadashi Matsunaga;Tomoko Yoshino
显示 I 类主要组织相容性复合物 (MHC I) 的磁性纳米粒子是使用趋磁细菌 Magnetospirillum 磁性 AMB-1 生物合成的。在细菌磁性颗粒（BacMP，也称为磁小体）上评估 MHC I 的表达水平和抗原肽 (HER263–71) 结合能力。此外，MHC I 和 HER263-71 的单链复合物是针对磁小体显示系统重新设计的，以改善 MHC I 和 HER263-71 之间的相互作用。测试了两种类型的融合排列，其中一种复合物估计以超过 70% 的水平折叠成正确的构象。除了高折叠率之外，该系统的一个优点是，即使 HER263-71 肽的 N 末端不是游离的，也不需要任何重折叠过程，而传统的细菌表达系统从未证明过这一点。所制备的单链 MHC I/HER263-71 复合物展示 BacMP（MHC I/HER2-BacMP）与 HER263-71 诱导的细胞特异性相互作用，并通过磁性分离，这表明天然 T 细胞受体可以识别 BacMP 上的工程化 MHC I/HER2 复合物。通过优化磁力分选方法，本研究中开发的 MHC I/HER2-BacMP 将在免疫治疗应用中发挥作用。
The magnetic nanoparticles displaying the class I major histocompatibility complex (MHC I) were biologically synthesized using the magnetotactic bacteriumMagnetospirillum magneticumAMB-1. Expression level and antigen peptide (HER263–71)-binding capability of the MHC I were evaluated on bacterial magnetic particles (BacMPs, also known as magnetosomes). Furthermore, the single-chain complexes of MHC I and HER263–71werede novodesigned for the magnetosome display system in order to improve the interaction between MHC I and HER263–71. Two types of the fusion arrangements were tested, and one of the complexes was estimated to fold into the correct conformation at the level of over 70%. In addition to the high folding ratio, an advantage of this system is that any refolding processes were not required even though theN-terminus of HER263–71peptide is not free, which conventional bacterial expression systems have never demonstrated. The as-prepared single-chain MHC I/HER263–71complex-displaying BacMPs (MHC I/HER2-BacMPs) specifically interacted with, and magnetically separated the HER263–71-induced cells, suggesting that the native T-cell receptor could recognize the engineered MHC I/HER2 complex on the BacMPs. By optimizing the magnetic sorting method, the MHC I/HER2-BacMPs developed in this study would be useful in immunotherapeutic applications.