OXYGEN DISTRIBUTION IN LIVING CELLS AS INVESTIGATED BY ESR MICROSCOPY

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 氧在动物细胞内的分布及其与低氧反应机制，特别是低氧诱导因子羟基酶的相互作用，是低氧生物学中一个重要的生物医学问题。斯科菲尔德教授的团队与牛津大学的拉特克利夫实验室和普格实验室(临床医学)合作，利用蛋白质组学、X射线结晶学、生物质谱学、分子生物学、动力学和有机合成等技术研究低氧反应的分子机制。在这里，我们将使用Acert的ESR显微镜设备，提供所研究的组织和细胞的微米分辨率，以分析在从低氧(2%)到常氧的范围内不同类型细胞中的氧分布。我们将通过结合细胞生物学和EPR引导的生物物理方法，将这些分析与感受酶的活性相关联。这些细胞将包括那些具有氧感应酶突变版本的细胞，这些酶在氧结合和反应方面具有不同的动力学特性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The distribution of oxygen within animal cells and how this interfaces with the hypoxia response machinery, especially the 'oxygen sensing' hypoxia inducible factor hydroxylases, is a major and biomedicinally important question in hypoxia biology. In collaboration with the Oxford-based Ratcliffe and Pugh laboratories (Clinical medicine) Professor Schofield's group studies the Molecular Mechanism of the Hypoxic Response using techniques such as proteomics, X-ray crystallography, biological mass spectrometry, molecular biology, kinetics and organic synthesis. Here we will use the ESR Microscopy facility at ACERT providing micrometer resolution of investigated tissues and cells to analyze oxygen distributions in different cell types under oxygen tensions ranging from hypoxia (2%) to normoxia. We will correlate these analyses with the activities of the sensing enzymes via a combined cell-biology and EPR led biophysical approaches. The cells will include those with mutant versions of the oxygen sensing enzymes that have varied kinetic properties with respect to oxygen binding and reaction.