利用SPR检测26肽与β1-肾上腺素受体自身抗体的相互作用

doi:10.3969/j.issn.1006-7795.2016.06.004

首都医科大学学报 ›› 2016, Vol. 37 ›› Issue (6): 736-739.doi: 10.3969/j.issn.1006-7795.2016.06.004

• 心血管疾病的病理生理机制 • 上一篇下一篇

利用SPR检测26肽与β₁-肾上腺素受体自身抗体的相互作用

董玉, 吕婷婷, 徐文丽, 白燕, 武烨, 刘慧荣, 王雯

首都医科大学基础医学院生理学与病理生理学系代谢紊乱相关心血管疾病北京市重点实验室, 北京 100069

收稿日期:2016-10-03 出版日期:2016-12-21 发布日期:2016-12-16
通讯作者: 王雯 E-mail:wangwen@ccmu.edu.cn
基金资助:
973计划前期研究专项（2014CB560704），北京市自然科学基金重点项目（7151001）

Detection of the interaction between the peptide and β₁-adrenergic receptor autoantibodies using surface plasmon resonance

Dong Yu, Lyu Tingting, Xu Wenli, Bai Yan, Wu Ye, Liu Huirong, Wang Wen

Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing Key Laboratory of Metabolic Disorders Related Cardiovascular Diseases, Capital Medical University, Beijing 100069, China

Received:2016-10-03 Online:2016-12-21 Published:2016-12-16
Supported by:
This study was supported by 973 Special Preliminary Study Plan(2014CB560704), Natural Science Foundation of Beijing(7151001).

摘要/Abstract

摘要： 目的探讨根据β₁肾上腺素受体细胞外第二环（β₁-adrenergic receptor，β₁-AR-EC_Ⅱ）的氨基酸序列合成的26肽与β₁-肾上腺素受体β₁-AR-EC_Ⅱ的单克隆抗体（β₁-adrenergic receptor autoantibodies，β₁-AA）的相互作用。方法根据人β₁-肾上腺素受体细胞外第二环（β₁-AR-EC_Ⅱ）的氨基酸序列合成26肽；采用杂交瘤细胞融合的方法获得针对β₁-AR-EC_Ⅱ的单克隆抗体（β₁-AA）；利用表面等离子共振（surface plasmon resonance，SPR）检测该合成的26肽与β₁-AA的亲和力；利用乳鼠心肌细胞跳动频率实验验证该合成26肽对β₁-AA的中和作用。结果提取的β₁-AA可以使乳鼠心肌细胞跳动频率增加（P<0.05），提示该β₁-AA具有生物学活性；合成26肽与β₁-AA结合的亲和力（K_D）为4.44 μmol/L，属于中等强度结合；较β₁-AA组相比，26肽处理后可明显降低心肌细胞跳动频率（P<0.05）。结论该合成26肽与β₁-肾上腺素受体自身抗体之间存在相互作用，并且可以拮抗β₁-AA引起的乳鼠心肌细胞跳动频率的增加。

关键词: β₁-肾上腺素受体自身抗体, 26肽, 表面等离子共振

Abstract: Objective To investigate the interaction between the peptide which mimic the structure of the second extracellular loop of the β₁-adrenergic receptor (β₁-AR-EC_Ⅱ) and β₁-adrenergic receptor autoantibodies (β₁-AA). Methods Peptide was synthesized according to the amino acid sequence of human β₁-AR-EC_Ⅱ; a hybridoma fusion method was used to obtain anti-β₁-AR-EC_Ⅱ monoclonal antibody; the interaction between the peptide and β₁-AA was detected by surface plasmon resonance (SPR); and the beating frequency experiment of neonatal rat cardiomyocytes was explored to test the neutralization of the peptide to β₁-AA. Results β₁-AA enhanced the beating frequency of neonatal rat cardiomyocytes (P<0.05), indicating that the β₁-AA we obtained has biological activity; SPR results showed that there was a moderate binding affinity (K_D) of 4.44 μmol/L between the synthetic peptide and β₁-AA. Conclusion There was interaction between the synthesized peptide and β₁-adrenergic receptor autoantibodies.

Key words: β₁-adrenergic receptor autoantibodies(β₁-AA), 26 peptide, surface plasmon resonance

中图分类号:

R971

董玉, 吕婷婷, 徐文丽, 白燕, 武烨, 刘慧荣, 王雯. 利用SPR检测26肽与β₁-肾上腺素受体自身抗体的相互作用[J]. 首都医科大学学报, 2016, 37(6): 736-739.

Dong Yu, Lyu Tingting, Xu Wenli, Bai Yan, Wu Ye, Liu Huirong, Wang Wen. Detection of the interaction between the peptide and β₁-adrenergic receptor autoantibodies using surface plasmon resonance[J]. Journal of Capital Medical University, 2016, 37(6): 736-739.

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利用SPR检测26肽与β₁-肾上腺素受体自身抗体的相互作用

Detection of the interaction between the peptide and β₁-adrenergic receptor autoantibodies using surface plasmon resonance

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