Glycolipid biointerface to decipher disease-implicated ganglioside-protein interactions

破译疾病相关神经节苷脂-蛋白质相互作用的糖脂生物界面 人体内的所有细胞，包括神经元、免疫细胞、上皮细胞和血细胞，都是 被一层厚厚的糖蛋白和糖脂包裹，被称为糖萼。不平凡的 糖萼的结构组织和生物合成的复杂性使其很难 理解它在各种细胞过程中扮演的确切角色，因此限制了它作为 治疗靶点。糖萼的一个重要分子家族是神经节苷脂，它参与 在广泛的细胞间事件中，如调节杀伤细胞毒性，控制神经再生， 并在炎症过程中促进细胞黏附。神经节苷脂被发现在 改变和介导膜蛋白在某些癌症中的亲和力特性，并明显 与胰岛素抵抗的2型糖尿病有关。然而，神经节苷脂的生化机制 对肿瘤和2型糖尿病的作用似乎极其复杂，神经节苷脂的主要成分 病理学仍然难以捉摸。缺乏合适的技术是主要限制 神经节苷脂的研究限制了我们了解它们在蛋白质功能中的作用的能力。 我们建议建立一种高效的、糖基多样性的仿生膜界面系统，并 一种新的生物分析平台研究神经节苷脂-蛋白质相互作用与几种疾病的关系 在分子水平上。将创建一个神经节苷脂文库，用于构建与 在这些疾病中观察到的精确控制的多糖部分、组成和包装生物物理 各州。所提出的方法绕过了神经节苷脂的复杂内源性合成，并创造了 一种新型的宿主环境，在神经节苷脂成分中进行程序化调节，以阐明结构- 与膜蛋白的功能关系。神经节苷脂对蛋白质相互作用的影响 主要通过表面等离子体共振(SPR)光谱进行研究，这种光谱可以量化分子 结合和亲和力在系统变化的组成和头基部分下的变化(目标1)。我们 然后将研究和了解神经节苷脂对EGFR和蛋白质的抑制/促进作用 VEGFR，与癌症进展相关的血管生成激活剂(目标2)，以及胰岛素的相互作用， 胰岛素受体和小窝蛋白-1(AIM 3)，一个与胰岛素抵抗的2型糖尿病有关的关键系统。

Glycolipid biointerface to decipher disease-implicated ganglioside-protein interactions All cells in the human body, including neurons, immune cells, epithelial cells, and blood cells, are coated with a dense layer of glycoproteins and glycolipids known as the glycocalyx. The extraordinary complexity in structural organization and biosynthesis of the glycocalyx has made it very difficult to comprehend the precise roles it plays in various cellular processes and thus limited its potential as therapeutic target. An important family of molecules of the glycocalyx is gangliosides, which participate in a wide array of intercellular events such as modulating killer cell toxicity, controlling neural regeneration, and promoting cell adhesion during inflammation. Gangliosides are found to play important roles in altering and mediating affinity properties of the membrane proteins in certain cancers, and are clearly implicated in insulin-resistant type 2 diabetes. However, the biochemical mechanisms of gangliosides’ effect on tumor and type 2 diabetes appear to be extremely complex, and a major portion of ganglioside pathology remains elusive. Lack of suitable techniques is a main obstacle that has principally limited the research on gangliosides and restricted our ability to understand their roles on protein function. We propose to build a highly effective, glyco-diverse, biomimetic membrane interface system and a new bioanalytical platform to study the ganglioside-protein interactions implicated in several diseases at the molecular level. A ganglioside library will be created for construction of interface mimics with precisely controlled glycan moiety, composition and packing biophysics as observed in those disease states. The proposed approach bypasses complex endogenous synthesis of gangliosides, and creates a novel hosting environment with programmed tuning in ganglioside makeups for elucidating structure- function relationships with the membrane proteins. The effect of gangliosides on protein interactions will be primarily investigated by surface plasmon resonance (SPR) spectroscopy, which quantifies molecular binding and affinity changes under systematically varied composition and headgroup moiety (Aim 1). We will then study and understand the inhibitory/promoting function of gangliosides on proteins EGFR and VEGFR, angiogenic activators linked to progression of cancer (Aim 2), and on interactions of insulin, insulin receptor and caveolin-1 (Aim 3), a key system implicated in insulin-resistant type 2 diabetes.