Development of solid-state NMR spectroscopic approaches for investigation of biological extracellular matrices

固态核磁共振是一种多用途的技术，可以提供处于功能相关状态的生物分子的结构信息。然而，单核磁共振在细胞外基质(ECM)生物材料中的应用仍然有限。细胞外基质种类繁多，通常由多种分子实体组成，包括蛋白质、多糖、糖蛋白和其他生物分子。这些分子在一定的长度范围内表现出特定的相互作用和组织。Ss核磁共振可以在原子长度尺度上提供结构洞察力，潜在地对体外样品(例如来自细胞培养的样品)和体外样品(例如来自患者活检组织的样品)进行最小程度的处理或纯化。本项目的目的是开发一种通用的方法来使用单核磁共振波谱来研究不同来源生物的细胞外基质。具体目标包括：开发使用单核磁共振结合快速MAS(60 kHz+)和高磁场的方法为细胞外基质带来新的洞察力。研究含层粘连蛋白的凝胶/涂层，层粘连蛋白是支持体外干细胞生长的关键糖蛋白研究从Ehler-Danlos综合征患者的哺乳动物成纤维细胞中研究细胞外基质。研究来自真菌裂殖酵母Pombe的细胞壁这个项目将建立在生物单核磁共振技术的基础上，用于新的应用，匹配以下主题和EPSRC职权范围内的主要兴趣：分析科学生物物理学医疗技术抗菌素耐药性(真菌)生物材料协作者包括华威医学院的Darius Koester博士和Prof Mohan Balasubramanian博士。

Solid-state nuclear magnetic resonance (ssNMR) is a versatile technique that can provide structural information on biomolecules in functionally-relevant states. However, applications of ssNMR to biological materials known as the extracellular matrix (ECM) remain limited. There is a diversity of ECMs, which are usually composed of a heterogeneous mixture of molecular entities, including proteins, glycans, glycoproteins, and other biomolecules. These molecules show specific interactions and organisation at a range of length scales. SsNMR can contribute structural insight at the atomic length scale, potentially on in vitro samples (e.g. from cell culture) and ex vivo samples (e.g. from patient biopsies) with minimal processing or purification.The aim of this project is to develop a general approach for investigating ECMs of different source organisms using ssNMR spectroscopy. Specific objectives include:Developing methods to bring new insight to ECMs using ssNMR combined with fast MAS (60 kHz+) and high magnetic fields.Investigation of gels/coatings containing laminin, a key glycoprotein used for supporting stem cell growth in vitroInvestigation of ECM from mammalian fibroblasts from patients with Ehlers-Danlos syndromes.Investigation of cell wall from the fungi Schizosaccharomyces pombeThis project will build on existing techniques in biological ssNMR for novel applications, matching the following topics and key interests within the remit of EPSRC:Analytical scienceBiophysicsHealthcare technologiesAntimicrobial resistance (fungal)BiomaterialsCollaborators for this project include Dr Darius Koester and Prof Mohan Balasubramanian of the Warwick Medical School.