Correlative cryo-microscopy: a new approach for characterizing the structure and
相关冷冻显微镜:一种表征结构和特征的新方法
基本信息
- 批准号:7904951
- 负责人:
- 金额:$ 33.56万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2008
- 资助国家:美国
- 起止时间:2008-08-01 至 2012-07-31
- 项目状态:已结题
- 来源:
- 关键词:BiologicalCell physiologyCellsComplexCryoelectron MicroscopyCrystallographyDevelopmentElectron MicroscopeFluorescenceGenerationsHuman poliovirusHybridsIceIn SituIn VitroLinkLocationMacromolecular ComplexesMapsMechanicsMethodsMicrofilamentsMicroscopyMicrotubulesModelingMolecularMolecular ChaperonesOpticsPathway interactionsPoliovirusesResolutionRibosomesStagingStructural BiologistStructureSubcellular structureTechniquesTechnologyViruscomputerized toolsimaging modalitymacromolecular assemblymulticatalytic endopeptidase complexnanometernew technologynovel strategiesoptical imagingreconstructionsingle moleculetomography
项目摘要
DESCRIPTION (provided by applicant): The macromolecular machines of cells pose structural questions that span over six orders of magnitude in scale from the tenths of nanometers (atoms) to tens of microns (cells). This large range of scale exceeds the capabilities of any one technique. As a result, structural biologists have increasingly turned to the use of hybrid approaches. For example, the combination of x-ray crystallography and cryoelectron microscopy has been widely used to probe the structure and mechanics of large complexes (e.g. viruses, ribosomes, proteosomes, chaperones, actin filaments and microtubules) in vitro at the near atomic level. In this project we will explore technologies to link these first generation hybrid techniques, single molecule optical microscopy, and cryoelectron tomography to characterize the structure and function of macromolecular complexes in the context of the whole cell. The technologies that will be developed include the use of optical imaging methods to characterize the functional state of macromolecular complexes in cells imbedded in vitreous ice and determine their location with very high precision (tens of nm), development of a cold stage that will permit the optical positional mapping to be transferred to the electron microscope, development of computational tools to locate, identify, and determine the structures of macromolecular complexes in cryoelectron tomographic reconstructions, and refinement of computational tools to match these structures with higher resolution structures derived from conventional in vitro methods. In our initial studies we will use the new technologies to characterize the cell entry pathway of poliovirus. This model was chosen because of its biological relevance (the cell entry mechanisms of nonenveloped viruses are poorly understood) and because structures of several relevant cell entry intermediates in vitro are already known. The techniques developed will be generally applicable to a wide range of macromolecular complexes.
In this project will develop technologies that will combine fluorescence optical microscopy and cryoelectron tomography to characterize the structure and function of intracellular macromolecular assemblies at the molecular level in situ. The technology will be relevant to a wide range of intracellular structures, and will add significantly to our basic understanding of the structure and function of "cellular machines."
描述(由申请人提供):细胞的大分子机器提出了跨越六个数量级的结构问题,从十分之一纳米(原子)到几十微米(细胞)。如此大范围的规模超出了任何一种技术的能力。因此,结构生物学家越来越多地转向使用混合方法。例如,x射线晶体学和低温电子显微镜的结合已被广泛用于在近原子水平上探测体外大型复合物(如病毒、核糖体、蛋白质体、伴侣蛋白、肌动蛋白丝和微管)的结构和力学。在这个项目中,我们将探索将这些第一代杂交技术、单分子光学显微镜和低温电子断层扫描技术联系起来的技术,以表征整个细胞背景下大分子复合物的结构和功能。将开发的技术包括使用光学成像方法来表征嵌入玻璃体冰的细胞中的大分子复合物的功能状态,并以非常高的精度(几十纳米)确定它们的位置,开发冷阶段,将允许光学位置映射转移到电子显微镜,开发计算工具来定位,识别,并在低温电子层析重建中确定大分子复合物的结构,并改进计算工具以使这些结构与传统体外方法获得的更高分辨率结构相匹配。在我们的初步研究中,我们将使用新技术来表征脊髓灰质炎病毒的细胞进入途径。选择该模型是因为其生物学相关性(非包膜病毒的细胞进入机制尚不清楚),并且因为几种相关的体外细胞进入中间体的结构已经已知。所开发的技术将普遍适用于广泛的大分子复合物。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
JAMES M HOGLE其他文献
JAMES M HOGLE的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('JAMES M HOGLE', 18)}}的其他基金
Correlative cryo-microscopy: a new approach for characterizing the structure and
相关冷冻显微镜:一种表征结构和特征的新方法
- 批准号:
8118885 - 财政年份:2008
- 资助金额:
$ 33.56万 - 项目类别:
Correlative cryo-microscopy: a new approach for characterizing the structure and
相关冷冻显微镜:一种表征结构和特征的新方法
- 批准号:
7514762 - 财政年份:2008
- 资助金额:
$ 33.56万 - 项目类别:
Correlative cryo-microscopy: a new approach for characterizing the structure and
相关冷冻显微镜:一种表征结构和特征的新方法
- 批准号:
7664279 - 财政年份:2008
- 资助金额:
$ 33.56万 - 项目类别:
Structual Analysis of Enterovirus Cell Entry Pathways
肠道病毒细胞进入途径的结构分析
- 批准号:
6437171 - 财政年份:2002
- 资助金额:
$ 33.56万 - 项目类别:
MACCHESS CONSORTIUM FOR LARGE MACROMOLECULAR STRUCTURES: HERPES VIRUS
MACCHESS 大分子结构联盟:疱疹病毒
- 批准号:
6667800 - 财政年份:2002
- 资助金额:
$ 33.56万 - 项目类别:
STRUCTURE OF ENZYME SUBSTRATE COMPLEX FOR GENERATION OF UDP N ACETYLMURAMIC ACID
产生 UDP N 乙酰胞壁酸的酶底物复合物的结构
- 批准号:
6586664 - 财政年份:2002
- 资助金额:
$ 33.56万 - 项目类别:
STRUCTURE OF ENZYME SUBSTRATE COMPLEX FOR GENERATION OF UDP N ACETYLMURAMIC ACID
产生 UDP N 乙酰胞壁酸的酶底物复合物的结构
- 批准号:
6658631 - 财政年份:2002
- 资助金额:
$ 33.56万 - 项目类别:
STRUCTURE OF ENZYME SUBSTRATE COMPLEX FOR GENERATION OF UDP N ACETYLMURAMIC ACID
产生 UDP N 乙酰胞壁酸的酶底物复合物的结构
- 批准号:
6437582 - 财政年份:2001
- 资助金额:
$ 33.56万 - 项目类别:
MACCHESS CONSORTIUM FOR LARGE MACROMOLECULAR STRUCTURES: HERPES VIRUS
MACCHESS 大分子结构联盟:疱疹病毒
- 批准号:
6491123 - 财政年份:2001
- 资助金额:
$ 33.56万 - 项目类别:
MACCHESS CONSORTIUM FOR LARGE MACROMOLECULAR STRUCTURES: HERPES VIRUS
MACCHESS 大分子结构联盟:疱疹病毒
- 批准号:
6339135 - 财政年份:2000
- 资助金额:
$ 33.56万 - 项目类别:
相似海外基金
Establishing the role of cell size dysregulation in cancer cell physiology and cellular ageing
确定细胞大小失调在癌细胞生理学和细胞衰老中的作用
- 批准号:
MR/X020290/1 - 财政年份:2024
- 资助金额:
$ 33.56万 - 项目类别:
Fellowship
Maestro Pro multiwell microelectrode array for the University of Liverpool electrophysiology suite: Cell physiology meets high throughput.
适用于利物浦大学电生理学套件的 Maestro Pro 多孔微电极阵列:细胞生理学满足高通量要求。
- 批准号:
BB/X019357/1 - 财政年份:2023
- 资助金额:
$ 33.56万 - 项目类别:
Research Grant
Investigating changes to marine organism excitable cell physiology following anthropogenic disturbances.
研究人为干扰后海洋生物可兴奋细胞生理学的变化。
- 批准号:
557505-2021 - 财政年份:2022
- 资助金额:
$ 33.56万 - 项目类别:
Postdoctoral Fellowships
CAREER: Investigating the Cellular Electrome as a Biomarker in Red Blood Cell Physiology and Pathology
职业:研究细胞电组作为红细胞生理学和病理学中的生物标志物
- 批准号:
2145313 - 财政年份:2022
- 资助金额:
$ 33.56万 - 项目类别:
Continuing Grant
Understanding the role of intracellular cholesterol transport in cell physiology
了解细胞内胆固醇转运在细胞生理学中的作用
- 批准号:
22H02620 - 财政年份:2022
- 资助金额:
$ 33.56万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Collaborative Research: Scaling from single-cell physiology to community stability in a natural gut microbiome
合作研究:从单细胞生理学扩展到天然肠道微生物群落的稳定性
- 批准号:
2032985 - 财政年份:2021
- 资助金额:
$ 33.56万 - 项目类别:
Continuing Grant
Investigating changes to marine organism excitable cell physiology following anthropogenic disturbances.
研究人为干扰后海洋生物可兴奋细胞生理学的变化。
- 批准号:
557505-2021 - 财政年份:2021
- 资助金额:
$ 33.56万 - 项目类别:
Postdoctoral Fellowships
Visualizing Live Cell Physiology with High Resolution Using Phase-Contrast STEM
使用相差 STEM 以高分辨率可视化活细胞生理学
- 批准号:
10224280 - 财政年份:2020
- 资助金额:
$ 33.56万 - 项目类别: