An integrated mass spectrometry approach to study heparin structure-bioactivity

研究肝素结构-生物活性的综合质谱方法

基本信息

批准号：
9252476
负责人：
IGOR A KALTASHOV
金额：
$ 31.19万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2020-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9252476
关键词：
Address Adopted Affinity Alzheimer&apos s Disease Anticoagulants Antithrombin III Area Binding Binding Sites Biopolymers Blood coagulation Cell Adhesion Cell Differentiation process Cell Proliferation Charge Chemistry Clinic Clinical Complex Crystallization Data Detection Deuterium Digestion Disease Dissociation Drug Delivery Systems Electrons Electrostatics Embryonic Development Exhibits Family Fibroblast Growth Factor Gases Glycosaminoglycans Goals Heparin Heparin Lyase Heparinoids Heterogeneity Hydrogen Inflammation Ion-Exchange Chromatography Procedure Ions Kinetics Knowledge Length Malignant Neoplasms Mass Spectrum Analysis Measures Medical Medicine Methodology Modality Modeling Molecular Molecular Conformation Molecular Models Monte Carlo Method Multiple Partners Neoplasm Metastasis Neurodegenerative Disorders Parkinson Disease Pathologic Pharmacology Phase Physiological Physiological Processes Polymers Polysaccharides Process Property Proteins Regenerative Medicine Research Resolution Role Source Spectrometry, Mass, Electrospray Ionization Structure Techniques Therapeutic Therapeutic Agents Therapeutic Effect Tissue Engineering Vertebral column Work Wound Healing angiogenesis base cationic antimicrobial protein CAP 37 clinical application design holistic approach immunogenic immunoregulation improved ion mobility member molecular modeling novel oncology protein complex public health relevance scaffold simulation stoichiometry structural biology therapeutic target tissue regeneration tool

项目摘要

DESCRIPTION (provided by applicant): Heparin-related glycosaminoglycans (GAGs) hold enormous promise in the field of regenerative medicine, and also are a largely unexploited source of therapeutic agents to address a range of other medically important conditions including cancer and neurodegenerative diseases. Nevertheless, their practical utilization in the clinic is still confined to anticoagulants, while progress in other areas remains relatively modest The great challenge presented by these biopolymers to medicine and, more broadly, to structural biology is due in large part to their enormous structural heterogeneity, which is mirrored by their multiple roles in modulating angiogenesis, cell adhesion, embryogenesis, inflammation, metastasis and wound healing. Despite extensive research efforts, clear understanding of how various structural features modulate function of these biopolymers remains wanting, arguably because the inquiry was designed using the framework of the lock-and-key model, a paradigm that had been tremendously successful in structural biology applied to proteins. However, this approach fails to recognize that the dramatically higher level of structural diversity exhibited by GAGs has functional importance. The proposed research seeks to shift the prevailing paradigm in the field of GAG/protein interactions by moving away from the notion of well-defined sterically complementary binding sites and emphasizing a greater role of polyvalent electrostatics. In addition to revealing the determinants of GAG/protein interactions and understanding how they fine-tune the binding process, we will obtain a high-resolution and dynamic description of the mechanism of heparin interaction with several therapeutically relevant proteins. This will be done without either limiting the structural space to a few precisel defined GAG molecules or reducing the conformational space to a few static snapshots available from crystal structures. Towards this goal, we will adopt a "holistic" approach to protein/GAG interactions that embraces the structural diversity of heparin-like GAGs, as opposed to the commonly accepted approach which limits the scope of inquiry to a few molecules accessible through synthesis. We will use a combination of bottom-up and top-down approaches to study protein/GAG interactions, the former focusing on relatively well-defined subsets of short heparin oligomers obtained by affinity separations, and the latter aimed at intact heparin. This will reveal the structural properties governing the interactions of GAGs with several therapeutically relevant proteins. Novel applications of gas-phase ion chemistry (electron capture and collisional activation) will be developed for characterization of large heterogeneous GAG/protein complexes. Finally, atomic-level Monte Carlo simulations in parallel with H/D exchange studies will lead to a high-resolution and dynamic depiction of the interaction process. This new view of GAG/protein interactions will allow the therapeutic potential of heparin-like GAGs to be exploited more efficiently in areas as diverse as regenerative medicine, inflammation, neurodegenerative disorders and oncology.

描述（由适用提供）：与肝素相关的糖胺聚糖（GAGS）在再生医学领域具有增强的希望，并且在很大程度上是意外的治疗剂来源，以解决其他医学上重要的疾病，包括癌症和神经退行性疾病。 Nevertheless, their practical utilization in the clinic is still confined to anticoagulants, while progress in other areas remain relatively modest The great challenge presented by these biopolymers to medicine and, more broadly, to structural biology is due in large part to their enormous structural heterogeneity, which is mirrored by their multiple roles in modulating angiogenesis, cell adhesion, embryogenesis, infection, metastasis and wound healing.尽管进行了广泛的研究工作，但对各种结构特征如何调节这些生物聚合物的功能的清晰了解，可以说是因为询问是使用锁定模型的框架设计的，该范围是在应用于蛋白质的结构生物学中取得了巨大成功的范式。但是，这种方法未能认识到，插科打s暴露的动态较高的结构多样性水平具有功能重要性。拟议的研究试图通过摆脱明确定义的空间互补结合位点的概念，并强调多价静电仪的作用，从而改变了GAG/蛋白质相互作用领域的现行范式。除了揭示插孔/蛋白质相互作用的决定剂并了解它们如何微调结合过程之外，我们还将获得对肝素相互作用机理与几种治疗相关蛋白质的高分辨率和动态描述。这将在不将结构空间限制为几个精液定义的GAG分子或将构象空间减少到晶体结构可用的一些静态快照的情况下进行。为了实现这一目标，我们将采用一种“整体”方法，用于蛋白质/插科打互动，该方法包含肝素状插科打的结构多样性，而不是将探究范围限制在通过合成中访问的一些分子的范围。我们将使用自下而上的方法和自上而下的方法来研究蛋白质/堵嘴的相互作用，前者着重于通过亲和力分离获得的短肝素低聚物的相对定义明确的子集，而后者则针对完整肝素。这将揭示控制插科打术与几种治疗相关蛋白质相互作用的结构特性。将开发出气相离子化学（电子捕获和碰撞激活）的新型应用，以表征大型异质GAG/蛋白质复合物。最后，与H/D交换研究并行的原子级蒙特卡洛模拟将导致相互作用过程的高分辨率和动态描述。这种GAG/蛋白质相互作用的新观点将使肝素样插科打s的治疗潜力在像再生医学，感染，神经退行性疾病和肿瘤学的潜水区中更有效地探索。