SPR and HDXMS analysis of interactions between thrombomodulin domain 1 and complement proteins
SPR and HDXMS analysis of interactions between thrombomodulin domain 1 and complement proteins
复制标题
SPR 和 HDXMS 分析血栓调节蛋白结构域 1 与补体蛋白之间的相互作用
DOI：
10.1016/j.bpj.2021.11.1844
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发表时间：
2022
影响因子：
3.4
通讯作者：
Scott, Michael
中科院分区：
文献类型：
--
作者：
Koeppe, Julia R.;Giler, Jose;Bodah, Timothy;Farrell, Kevin;Gomez, Chelsea;Scott, Michael
我们使用表面等离子共振（SPR）和氢/氘交换质谱（HDXMS）来研究血栓调节蛋白和补体蛋白（包括补体成分3（C3）和补体因子H（CFH））之间的相互作用。补体成分 3 (C3) 位于三种不同补体激活途径（经典、凝集素和替代）的交汇处。 C3b 的激活是先天免疫反应的关键步骤，可形成重要的多蛋白复合物，最终参与病原体清除。当补体调节不当时，会导致炎症性疾病和自身免疫性疾病。 CFH 是 C3b 的几种调节蛋白之一，然而，补体系统中调节相互作用的完整分子细节尚未阐明。血栓调节蛋白 (TM)，特别是其 N 末端凝集素样结构域 (TMD1)，已被确定为通过与 C3 或 C3b 相互作用的可能的补体调节剂，并且可能还需要已知的调节剂 CFH。使用 SPR，我们发现 C3 或 C3b 可以结合到涂有 TMD1 的表面，并且完整的动力学研究正在进行中。我们还使用 SPR 研究 TMD1 和 CFH 之间的相互作用。使用 HDXMS，我们看到 TMD1 与 C3b 相互作用，与 C3 的相互作用较少。 TMD1 倾向于使 C3b 更容易进行氘交换，而在 TMD1 存在的情况下，C3 往往不太容易进行氘交换。这种差异表明 TMD1 在调节补体途径的关键步骤中可能发挥作用。
We have used surface plasmon resonance (SPR) and hydrogen/deuterium exchange mass spectrometry (HDXMS) to study the interaction between thrombomodulin and complement proteins including complement component 3 (C3) and complement factor H (CFH). Complement component 3 (C3) is at the junction of three different complement activation pathways (classical, lectin, and alternative). Activation to C3b is a key step in the innate immune response that allows for the formation of important multi-protein complexes that ultimately participate in pathogen clearance. When misregulated, complement can lead to inflammatory disease and autoimmune disorders. CFH is one of several regulatory proteins for C3b, however, the full molecular details of the regulatory interactions in the complement system have not yet been elucidated. Thrombomodulin (TM), and specifically its N-terminal lectin-like domain (TMD1), has been identified as a possible regulator of complement through interactions with C3 or C3b, and the known regulator CFH may also be required. Using SPR, we see that C3 or C3b can bind to a surface coated with TMD1, and full kinetic studies are underway. We are also investigating interactions between TMD1 and CFH using SPR. Using HDXMS, we see that TMD1 interacts with C3b, and there is a lesser interaction with C3. TMD1 tends to make C3b more accessible to deuterium exchange, while C3 tends to be less accessible to deuterium exchange in the presence of TMD1. This difference suggests a possible role for TMD1 in regulating a key step in the complement pathway.