Department of Chemistry, Physical and Theoretical Chemistry Laboratory, University of Oxford, Oxford, UK; The Kavli Institute for Nanoscience Discovery, Dorothy Crowfoot Hodgkin Building, University of Oxford, Oxford, UK; King's College London British Heart Foundation Centre, School of Cardiovascular Medicine and Sciences, London, UK.
Department of Infectious Diseases, School of Immunology and Microbial Sciences, King's College London, London, UK.
Mol Cell Proteomics. 2023 Aug;22(8):100600. doi: 10.1016/j.mcpro.2023.100600. Epub 2023 Jun 19.
High-density lipoprotein (HDL) levels are reduced in patients with coronavirus disease 2019 (COVID-19), and the extent of this reduction is associated with poor clinical outcomes. While lipoproteins are known to play a key role during the life cycle of the hepatitis C virus, their influence on coronavirus (CoV) infections is poorly understood. In this study, we utilize cross-linking mass spectrometry (XL-MS) to determine circulating protein interactors of the severe acute respiratory syndrome (SARS)-CoV-2 spike glycoprotein. XL-MS of plasma isolated from patients with COVID-19 uncovered HDL protein interaction networks, dominated by acute-phase serum amyloid proteins, whereby serum amyloid A2 was shown to bind to apolipoprotein (Apo) D. XL-MS on isolated HDL confirmed ApoD to interact with SARS-CoV-2 spike but not SARS-CoV-1 spike. Other direct interactions of SARS-CoV-2 spike upon HDL included ApoA1 and ApoC3. The interaction between ApoD and spike was further validated in cells using immunoprecipitation-MS, which uncovered a novel interaction between both ApoD and spike with membrane-associated progesterone receptor component 1. Mechanistically, XL-MS coupled with data-driven structural modeling determined that ApoD may interact within the receptor-binding domain of the spike. However, ApoD overexpression in multiple cell-based assays had no effect upon viral replication or infectivity. Thus, SARS-CoV-2 spike can bind to apolipoproteins on HDL, but these interactions do not appear to alter infectivity.
高密度脂蛋白（HDL）水平在 2019 年冠状病毒病（COVID-19）患者中降低，并且这种降低的程度与不良的临床结局相关。虽然脂蛋白在丙型肝炎病毒的生命周期中起着关键作用，但它们对冠状病毒（CoV）感染的影响知之甚少。在这项研究中，我们利用交联质谱（XL-MS）来确定严重急性呼吸综合征（SARS）-CoV-2 刺突糖蛋白的循环蛋白相互作用体。从 COVID-19 患者血浆中分离出的 XL-MS 揭示了 HDL 蛋白相互作用网络，主要由急性期血清淀粉样蛋白组成，其中血清淀粉样蛋白 A2 被证明与载脂蛋白（Apo）D 结合。在分离的 HDL 上进行的 XL-MS 证实 ApoD 与 SARS-CoV-2 刺突而非 SARS-CoV-1 刺突相互作用。SARS-CoV-2 刺突与 HDL 之间的其他直接相互作用包括 ApoA1 和 ApoC3。使用免疫沉淀-MS 在细胞中进一步验证了 ApoD 与刺突之间的相互作用，该方法揭示了 ApoD 与刺突以及膜相关孕激素受体成分 1 之间的新相互作用。从机制上讲，XL-MS 与数据驱动的结构建模相结合，确定 ApoD 可能在刺突的受体结合域内相互作用。然而，在多种基于细胞的测定中过表达 ApoD 对病毒复制或感染性没有影响。因此，SARS-CoV-2 刺突可以与 HDL 上的载脂蛋白结合，但这些相互作用似乎不会改变感染性。
