THE PERIPHERAL STALK OF YEAST VACUOLAR ATPASE

酵母液泡ATP酶的外周柄

基本信息

  • 批准号:
    8363542
  • 负责人:
  • 金额:
    $ 1.78万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2011
  • 资助国家:
    美国
  • 起止时间:
    2011-07-01 至 2012-06-30
  • 项目状态:
    已结题

项目摘要

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 vacuolar ATPase (V-ATPase) is a multi subunit rotary motor enzyme that functions as an ATP hydrolysis driven proton pump in the endomembrane system of eukaryotic cells. The V-ATPase consistes of two motor domains, a cytoplasmic ATPase (V1) and a membrane bound proton channel (V0). The two domains are linked by three peripheral stator proteins that function as a structural link to counteract the rotational torque that is generated during ATP hydrolysis. We have recently obtained a 3-D reconstruction of the intact V-ATPase from yeast (Zhang et al., JBC 283, 35983) and we are now using X-ray crystallography to determine the atomic resolution structures of V-ATPase subunits and subunit domains for fitting into the EM derived map. We have crystallized the peripheral stalk forming subunits of yeast V-ATPase (subunits E&G) in complex with a domain of subunit C. A preliminary diffraction analysis performed at the Chess beamline F1 (Fall 2009; in collaboration with Dr. Edward Berry) resulted in ~5.5 ¿ diffraction. The crystals belong to spacegroup P212121 with unit cell parameters of 95.2, 114.1, 133.9 ¿ , 90,90.5,90¿¿. Currently, there is no crystal structure available for the peripheral stalk(s) of the V-ATPase (or any of the related rotary ATPases including the F1F0-ATP synthase or the archaeal A-ATPase). SInce last fall, we have optimized crystallization conditions including a 96 condition additive screen, leading to numerous conditions with varying crystal morphologies. We will use the beam time at Chess, if approved, to screen crystals for high quality diffraction and to collect native data sets if time and crystal quality permits. Subsequent work, for which a full proposal is planned, would include heavy metal soaks and/or SeMet containing protein. Molecular replacement may be possible as a crystal structure for subunit C is available.
这个子项目是利用这些资源的众多研究子项目之一

项目成果

期刊论文数量(0)
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科研奖励数量(0)
会议论文数量(0)
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Stephan Wilkens其他文献

Stephan Wilkens的其他文献

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{{ truncateString('Stephan Wilkens', 18)}}的其他基金

Structure and Regulatory Mechanisms of the Vacuolar ATPase
液泡ATP酶的结构和调节机制
  • 批准号:
    10612863
  • 财政年份:
    2021
  • 资助金额:
    $ 1.78万
  • 项目类别:
A freezing robot for cryo-EM
用于冷冻电镜的冷冻机器人
  • 批准号:
    10581952
  • 财政年份:
    2021
  • 资助金额:
    $ 1.78万
  • 项目类别:
Structure and Regulatory Mechanisms of the Vacuolar ATPase
液泡ATP酶的结构和调节机制
  • 批准号:
    10206746
  • 财政年份:
    2021
  • 资助金额:
    $ 1.78万
  • 项目类别:
Structure and Regulatory Mechanisms of the Vacuolar ATPase
液泡ATP酶的结构和调节机制
  • 批准号:
    10398935
  • 财政年份:
    2021
  • 资助金额:
    $ 1.78万
  • 项目类别:
A novel tool for organelle and isoform specific targeting of V-ATPase in cancer
癌症中 V-ATP 酶的细胞器和亚型特异性靶向的新工具
  • 批准号:
    9764745
  • 财政年份:
    2019
  • 资助金额:
    $ 1.78万
  • 项目类别:
A 800 MHz Nuclear Magnetic Resonance Spectrometer in Support of Life Science Rese
支持生命科学研究的 800 MHz 核磁共振波谱仪
  • 批准号:
    8334960
  • 财政年份:
    2013
  • 资助金额:
    $ 1.78万
  • 项目类别:
THE PERIPHERAL STALK OF YEAST VACUOLAR ATPASE
酵母液泡ATP酶的外周柄
  • 批准号:
    8171534
  • 财政年份:
    2010
  • 资助金额:
    $ 1.78万
  • 项目类别:
STRUCTURE OF THE VACUOLAR ATPase
液泡ATP酶的结构
  • 批准号:
    8000056
  • 财政年份:
    2010
  • 资助金额:
    $ 1.78万
  • 项目类别:
STRUCTURE OF P-GLYCOPROTEIN BY ELECTRON MICROSCOPY
电子显微镜下 P-糖蛋白的结构
  • 批准号:
    7092555
  • 财政年份:
    2006
  • 资助金额:
    $ 1.78万
  • 项目类别:
STRUCTURE OF P-GLYCOPROTEIN BY ELECTRON MICROSCOPY
电子显微镜下 P-糖蛋白的结构
  • 批准号:
    7408550
  • 财政年份:
    2006
  • 资助金额:
    $ 1.78万
  • 项目类别:

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