Acquisition of a 700 MHz NMR CryoProbe

摘要 这项提议的首要目标是了解分子运动和生物物理性质 调节蛋白质的相互作用，以促进正常的动态平衡或病理疾病状态。我们在调查 三种情况下蛋白质动力学与相互作用的关系：1)蛋白质-纤维状胶原 参与血小板聚集的相互作用；2)α-突触核蛋白(αS)，一种固有的无序蛋白 其错误折叠和聚集成淀粉样纤维和沉积到路易氏小体与 衰弱的突触核病症，如帕金森氏病；以及3)严重急性 呼吸综合征冠状病毒2型，其与血管紧张素转换酶2的相互作用 (ACE2)通过受体结合域(RBD)是进入宿主细胞的起始点。我们最近做了 发现在每一种情况下，构象动力学都深刻地影响着它们从原子到纳米的尺度 并可能影响其生物分子相互作用的潜力。尽管在生物学上具有重要的意义 这些系统及其在疾病上的相互作用的含义，这些系统的分子决定因素 蛋白质之间的相互作用仍然没有答案。因此，我们使用多方面的方法来集成解决方案 以及带有生物物理、生物和计算方法的固态核磁共振，以解决分子 运动调节蛋白质的相互作用，以促进正常的动态平衡或病理疾病状态。收获 蛋白质与这些动态系统相互作用的分子理解依赖于状态- 最先进的解决方案核磁共振仪器能够准确地测量这些 用于检测相互作用事件并检测短暂的低密度复合体。

Abstract The overarching goal in this proposal is to understand how molecular motions and biophysical properties modulate protein interactions to promote normal homeostasis or pathological disease states. We investigate the relationship between protein dynamics and interactions in three contexts: 1) protein–fibrillar collagen interactions involved in platelet aggregation; 2) α-synuclein (αS), an intrinsically disordered protein (IDP) whose misfolding and aggregation into amyloid fibrils and deposition into Lewy bodies are associated with debilitating synucleinopathies, such as Parkinson's Disease; and 3) the spike glycoprotein of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), whose interaction with angiotensin converting enzyme 2 (ACE2) via the receptor binding domain (RBD) is the initial point of host cell entry. We have recently discovered that in each of these cases, conformational dynamics profoundly impact their atomic-to-nano scale properties and may affect their potential for biomolecular interactions. Despite the biological importance of these systems and the implications for their interactions on disease, the molecular determinants of these protein–protein interactions remain unanswered. Thus, we use a multifaceted approach integrating solution and solid-state NMR with biophysical, biological, and computational methods to address how molecular motions modulate protein interactions to promote normal homeostasis or pathological disease states. Gaining a molecular understanding of protein–protein interactions with each of these dynamic systems relies on state- of-the-art solution NMR instrumentation to be able to accurately measure timescales and fluctuations of these interaction events and to detect transient, lowly populated complexes.