Macrocarpal B blocks the binding between the phospholipase A2 receptor and its antibodies

IF 3.3 3区 医学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Zixin Feng , Fu-sheng Guo , Qian Wang , Miao Wang , Ming-Hui Zhao , Zhao Cui , Xiaoguang Lei
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Abstract

The pathogenic role of anti-phospholipase A2 receptor (PLA2R) antibodies in primary membranous nephropathy (MN) has been well-established. This study aimed to identify potential small-molecule inhibitors against the PLA2R-antibody interaction, offering potential therapeutic benefits. A comprehensive screening of over 4000 small-molecule compounds was conducted by ELISA to assess their inhibitory effects on the binding between the immobilized full-length extracellular PLA2R and its antibodies. The affinity of anti-PLA2R IgG from MN patients and the inhibitory efficacy of each compound were evaluated via surface plasmon resonance (SPR). Human podocyte injuries were analyzed using CCK-8 assay, wound healing assay, western blot analysis, and immunofluorescence, after exposure to MN plasma +/- blocking compound. Fifteen compounds were identified as potential inhibitors, demonstrating inhibition rates >20 % for the PLA2R-antibody interaction. Anti-PLA2R IgG exhibited a consistent affinity among patients (KD = 10−8 M). Macrocarpal B emerged as the most potent inhibitor, reducing the antigen–antibody interaction by nearly 30 % in a dose-dependent manner, comparable to the performance of the 31-mer peptide from the CysR domain. Macrocarpal B bound to the immobilized PLA2R with an affinity of 1.47 × 10−6 M, while showing no binding to anti-PLA2R IgG. Human podocytes exposed to MN plasma showed decreased podocin expression, impaired migration function, and reduced cell viability. Macrocarpal B inhibited the binding of anti-PLA2R IgG to podocytes and reduced the cellular injuries.

Abstract Image

大果桉醛 B 可阻断磷脂酶 A2 受体与其抗体之间的结合。
抗磷脂酶 A2 受体(PLA2R)抗体在原发性膜性肾病(MN)中的致病作用已得到证实。本研究旨在确定潜在的小分子PLA2R-抗体相互作用抑制剂,从而提供潜在的治疗益处。研究人员通过酶联免疫吸附试验(ELISA)对 4000 多种小分子化合物进行了全面筛选,以评估它们对固定的全长细胞外 PLA2R 与其抗体之间结合的抑制作用。通过表面等离子体共振(SPR)评估了来自 MN 患者的抗 PLA2R IgG 的亲和力和每种化合物的抑制效果。在暴露于 MN 血浆 +/- 阻断化合物后,使用 CCK-8 检测法、伤口愈合检测法、Western 印迹分析法和免疫荧光法分析了人类荚膜细胞损伤情况。15种化合物被确定为潜在的抑制剂,对PLA2R-抗体相互作用的抑制率大于20%。不同患者的抗 PLA2R IgG 具有一致的亲和力(KD = 10-8 M)。大果桉醛 B 是最有效的抑制剂,它以剂量依赖的方式将抗原-抗体相互作用降低了近 30%,与 CysR 结构域的 31-mer 肽的效果相当。大果桉醛 B 与固定的 PLA2R 的亲和力为 1.47 × 10-6 M,而与抗 PLA2R IgG 没有结合。暴露于 MN 血浆的人类荚膜细胞显示出荚膜蛋白表达减少、迁移功能受损和细胞活力降低。大果桉醛 B 可抑制抗-PLA2R IgG 与荚膜细胞的结合,减轻细胞损伤。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Bioorganic & Medicinal Chemistry
Bioorganic & Medicinal Chemistry 医学-生化与分子生物学
CiteScore
6.80
自引率
2.90%
发文量
413
审稿时长
17 days
期刊介绍: Bioorganic & Medicinal Chemistry provides an international forum for the publication of full original research papers and critical reviews on molecular interactions in key biological targets such as receptors, channels, enzymes, nucleotides, lipids and saccharides. The aim of the journal is to promote a better understanding at the molecular level of life processes, and living organisms, as well as the interaction of these with chemical agents. A special feature will be that colour illustrations will be reproduced at no charge to the author, provided that the Editor agrees that colour is essential to the information content of the illustration in question.
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