Division of Biotechnology and Life Science, Institute of Engineering, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
Int J Mol Sci. 2013 Jul 11;14(7):14426-38. doi: 10.3390/ijms140714426.
The measurement of autoantibodies to thyroid-stimulating hormone receptor (TSHR) is important for the diagnosis of autoimmune thyroid disease such as Graves' disease (GD). Although TSHR from porcine thyroid membrane is commonly used for the measurement of TSHR autoantibodies (TRAb), recombinant human TSHR (hTSHR) remains ideal in terms of stable supply and species identity. Here we set out to express recombinant hTSHR on the lipid-bilayer surface of magnetic nanoparticles from a magnetotactic bacterium, Magnetospirillum magneticum AMB-1. Using a tetracycline-inducible expression system, we successfully overexpressed functional hTSHR on bacterial magnetic particles (BacMPs) in AMB-1 via an anchor protein specific for BacMPs. The overexpressed hTSHR was membrane integrated and possessed both ligand and autoantibody binding activity. Our data suggest that hTSHR-displayed BacMPs have potential as novel tools for ligand-receptor interaction analysis or for TRAb immunoassay in GD patients.
测定甲状腺刺激激素受体(TSHR)自身抗体对于诊断自身免疫性甲状腺疾病(如格雷夫斯病(GD))非常重要。虽然猪甲状腺膜来源的 TSHR 常用于 TSHR 自身抗体(TRAb)的测定,但从稳定供应和物种同源性的角度来看,重组人 TSHR(hTSHR)仍然是理想的选择。在这里,我们旨在从趋磁细菌 Magnetospirillum magneticum AMB-1 的脂质双层表面表达重组 hTSHR。我们使用四环素诱导表达系统,通过针对 BacMPs 的锚定蛋白在 AMB-1 中的细菌磁珠(BacMPs)上成功过表达功能 hTSHR。过表达的 hTSHR 整合到膜上,具有配体和自身抗体结合活性。我们的数据表明,hTSHR 展示的 BacMPs 具有作为新型工具用于配体-受体相互作用分析或 GD 患者 TRAb 免疫测定的潜力。