Specific domain V reduction of beta-2-glycoprotein I induces protein flexibility and alters pathogenic antibody binding.
Specific domain V reduction of beta-2-glycoprotein I induces protein flexibility and alters pathogenic antibody binding.
复制标题
DOI：
10.1038/s41598-021-84021-2
复制
发表时间：
2021-02-25
影响因子：
4.6
通讯作者：
Delcea M
中科院分区：
文献类型：
--
作者：
Buchholz I;McDonnell T;Nestler P;Tharad S;Kulke M;Radziszewska A;Ripoll VM;Schmidt F;Hammer E;Toca-Herrera JL;Rahman A;Delcea M
β -2-糖蛋白I （β2GPI）是一种血液蛋白，是自身免疫性疾病抗磷脂综合征（APS）的主要抗原。β2GPI主要以封闭或开放构象存在，由分布在5个结构域的11个二硫化物组成。在结构域V的末端Cys288/Cys326二硫键与不同的半胱氨酸氧化还原态有关。这种二硫键在蛋白质构象动力学中的作用至今尚未被研究。在这里，我们报道了硫氧还蛋白-1（由Tris（2-羧基乙基）膦回收）的酶促还原；TCEP) β2GPI。Western blot和质谱分析证实了特异性的减少，证实了大多数靶向β2GPI的第五结构域。原子力显微镜图像显示，与模型研究证实的未经处理的蛋白质相比，减少的β2GPI显示出稍高的开放构象比例，并且更灵活。我们已经通过ELISA证实了致病性APS自身抗体与降低的β2GPI结合的强烈增加。我们的研究有助于了解β2GPI减少对APS患者蛋白结构的影响及其对抗体结合的影响。
Beta-2-glycoprotein I (β2GPI) is a blood protein and the major antigen in the autoimmune disorder antiphospholipid syndrome (APS). β2GPI exists mainly in closed or open conformations and comprises of 11 disulfides distributed across five domains. The terminal Cys288/Cys326 disulfide bond at domain V has been associated with different cysteine redox states. The role of this disulfide bond in conformational dynamics of this protein has not been investigated so far. Here, we report on the enzymatic driven reduction by thioredoxin-1 (recycled by Tris(2-carboxyethyl)phosphine; TCEP) of β2GPI. Specific reduction was demonstrated by Western blot and mass spectrometry analyses confirming majority targeting to the fifth domain of β2GPI. Atomic force microscopy images suggested that reduced β2GPI shows a slightly higher proportion of open conformation and is more flexible compared to the untreated protein as confirmed by modelling studies. We have determined a strong increase in the binding of pathogenic APS autoantibodies to reduced β2GPI as demonstrated by ELISA. Our study is relevant for understanding the effect of β2GPI reduction on the protein structure and its implications for antibody binding in APS patients.
登录
查看更多内容
影响因子：
13.3
作者：
Duarte-Garcia, Ali;Pham, Michael M.;Matteson, Eric. L.
通讯作者：
Matteson, Eric. L.
影响因子：
11.4
作者：
Bouma, B;de Groot, PG;Gros, P
通讯作者：
Gros, P
DOI：
10.1016/j.bbapap.2019.03.007
发表时间：
2019-06-01
影响因子：
3.2
作者：
Baerenfaenger, Melissa;Meyer, Bernd
通讯作者：
Meyer, Bernd
影响因子：
5
作者：
de Laat, Bas;de Groot, Philip G.
通讯作者：
de Groot, Philip G.
影响因子：
4.4
作者：
JORGENSEN, WL;CHANDRASEKHAR, J;KLEIN, ML
通讯作者：
KLEIN, ML