Smorodintsev Research Institute of Influenza, Russian Ministry of Health, 15/17 Ulitsa Prof. Popova, St. Petersburg, Russia, 197376.
Peter the Great Saint Petersburg Polytechnic University, 29 Ulitsa Polytechnicheskaya, St. Petersburg, Russia, 194064.
Biometals. 2022 Dec;35(6):1157-1168. doi: 10.1007/s10534-022-00432-2. Epub 2022 Aug 13.
The work is devoted to the study of the structural characteristics of the myeloperoxidase-ceruloplasmin-thrombin complex using small-angle neutron scattering methods in combination with computer modeling, as well as surface plasmon resonance and solid-phase enzyme assay. We have previously shown that the functioning of active myeloperoxidase during inflammation, despite the presence in the blood of an excess of ceruloplasmin which inhibits its activity, is possible due to the partial proteolysis of ceruloplasmin by thrombin. In this study, the myeloperoxidase-ceruloplasmin-thrombin heterohexamer was obtained in vitro. The building of a heterohexamer full-atomic model in silico, considering the glycosylation of the constituent proteins, confirmed the absence of steric barriers for the formation of protein-protein contacts. It was shown that the partial proteolysis of ceruloplasmin does not affect its ability to bind to myeloperoxidase, and a structural model of the heterohexamer was obtained using the small-angle neutron scattering method.
这项工作致力于使用小角中子散射方法结合计算机建模，以及表面等离子体共振和固相酶测定法，研究髓过氧化物酶-铜蓝蛋白-凝血酶复合物的结构特征。我们之前已经表明，尽管血液中存在过量的抑制其活性的铜蓝蛋白，但在炎症期间活性髓过氧化物酶的功能是可能的，这是由于凝血酶对铜蓝蛋白的部分蛋白水解。在这项研究中，体外获得了髓过氧化物酶-铜蓝蛋白-凝血酶异六聚体。在考虑到组成蛋白质的糖基化的情况下，在计算机上构建异六聚体全原子模型，证实了形成蛋白质-蛋白质接触不存在空间位阻。结果表明，铜蓝蛋白的部分蛋白水解并不影响其与髓过氧化物酶结合的能力，并且使用小角中子散射方法获得了异六聚体的结构模型。
