Super-resolution imaging of protein dynamics in the extracellular matrix

细胞外基质中蛋白质动力学的超分辨率成像

• 批准号: 10271827
• 负责人: Lydia Kisley
• 金额: $ 35.37万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10271827
• 关键词: Affect; Area; Biochemical; Biological; Biological Process; Biology; Biophysics; Cells; Chemicals; Complex; Development; Diffusion; Disease; Drug Delivery Systems; Environment; Extracellular Matrix; Extracellular Matrix Proteins; Goals; Image; Methods; Microscopy; Optics; Property; Protein Dynamics; Proteins; Regulation; Research Personnel; Resolution; Signaling Protein; Structure; Techniques; Therapeutic; Time; Tissue Engineering; Tissues; Variant; base; extracellular; imaging modality; model design; nanoscale; protein folding; protein function; spatiotemporal; temporal measurement

Project Summary We seek to understand how the physiochemical environment of the extracellular matrix affects the diffusion, structure, and folding of proteins. The extracellular matrix is spatially heterogeneous at nanoscales and can change over time. But current imaging methods are limited in spatial and temporal resolutions, while biochemical techniques are designed for model, pristine solutions that differ from the complexity of the extracellular matrix. Therefore, how proteins function locally in the extracellular matrix is an underexplored area in biophysics. We propose to understand how proteins function and interact in the complex extracellular matrix environment, and how the nanoscale variation in the chemical and physical composition of this environment can locally change protein dynamics. To achieve this, we will use a correlation-based super-resolution microscopy developed by our lab that can access the time- and spatial-scales of protein dynamics within the extracellular matrix. The goals of this proposal are to: 1) identify how the nanoscale chemical and steric properties of the extracellular matrix control protein diffusion, structure, and folding; 2) determine how cells tune the dynamics of proteins by changing the extracellular matrix; 3) develop instrumental and analysis methods that access the spatiotemporal regimes required to understand protein function in the extracellular environment. Achieving our goals will reveal the extracellular matrix and protein dynamics that drive biological regulation and will provide researchers with methods to understand and potentially control extracellular delivery of signaling proteins or therapeutics. Overall, an entirely new area of biology – the extracellular matrix - will be explored by super-resolution microscopy; the findings can be broadly applied to other biomolecules, cells, and tissues of relevance to fundamental biophysics, drug-delivery/therapeutics, and disease.

项目摘要 我们试图了解细胞外基质的理化环境如何影响扩散， 蛋白质的结构和折叠。细胞外基质在纳米尺度上是空间异质的， 随着时间的推移而改变。但目前的成像方法在空间和时间分辨率上受到限制，而生物化学成像方法则是一种新的成像方法。 技术是为模型而设计的，原始的解决方案不同于细胞外基质的复杂性。 因此，蛋白质如何在细胞外基质中局部发挥作用是生物物理学中尚未充分研究的领域。我们 建议了解蛋白质如何在复杂的细胞外基质环境中发挥作用和相互作用， 环境中化学和物理成分的纳米级变化如何局部改变 蛋白质动力学为了实现这一目标，我们将使用由美国国家航空航天局开发的基于相关性的超分辨率显微镜。 我们的实验室可以访问细胞外基质内蛋白质动力学的时间和空间尺度。的目标 1）确定细胞外基质的纳米级化学和空间性质如何 控制蛋白质的扩散、结构和折叠; 2）确定细胞如何通过以下方式调节蛋白质的动力学： 改变细胞外基质; 3）开发工具和分析方法， 了解蛋白质在细胞外环境中的功能所需的制度。实现我们的目标将揭示 细胞外基质和蛋白质动力学驱动生物调节，并将为研究人员提供 理解和潜在地控制信号蛋白或治疗剂的细胞外递送的方法。总的来说， 一个全新的生物学领域-细胞外基质-将通过超分辨率显微镜进行探索; 这些发现可以广泛地应用于与基础生物物理学相关的其他生物分子、细胞和组织， 药物递送/治疗和疾病。