CAREER: Conformational Changes in Voltage-Controlled Ion Channels Measured by Advanced Fluorescence Techniques
职业:通过先进荧光技术测量压控离子通道的构象变化
基本信息
- 批准号:9984841
- 负责人:
- 金额:$ 50万
- 依托单位:
- 依托单位国家:美国
- 项目类别:Continuing Grant
- 财政年份:2000
- 资助国家:美国
- 起止时间:2000-04-15 至 2006-03-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
This project describes an interdisciplinary five-year career development plan involving physics, chemistry, and biology in both research and teaching.In research, two complementary fluorescence techniques will be used to study conformational changes in ion channels-channels that are responsible for nerve conduction. These techniques, one of which measures distance and the other of which measures rotation, can be applied to many other kinds of proteins and channels. The first technique, lanthanide-based resonance energy transfer (LRET), is an improved version of an established technique called fluorescence resonance energy transfer. This improved resolution is achieved because the lanthanide can transfer energy to an organic dye by a mechanism that is sensitive to angstrom changes in distance between the two labels. The second method is a version of phosphorescence anisotropy utilizing either phosphorescent probes or our recently discovered polarized luminescent lanthanide probes. In this technique, a polarized excitation pulse selectively excites those probes aligned with the pulse polarization and polarization-sensitive detection monitors the reorientation of probes. The technique has the advantage that it can measure rotational motion on the millisecond time scale, the time-scale associated with protein rearrangements in ion channels. LRET will be used to measure distances between site-specifically placed lanthanide atoms and organic dyes as a function of voltage across the cell membrane and relate them to motion within the channel during simultaneous electrical measurements. Measuring rotation of appropriately placed probes by phosphorescence anisotropy can further characterize protein movement. Initial studies with LRET have achieved a milestone in the field of voltage-controlled ion channels, the first measurements of actual distance changes as ion channel proteins move in response to voltage changes. These results suggest that part of the channel rotates, a hypothesis that will be tested further. In teaching, an undergraduate course intended primarily for non-scientists, entitled "Science and the Citizen: Current controversies in science, technology and society" will be developed. The course will be taught in a "pro-con" format with introductory lectures giving students background in contemporary science issues, followed by readings from advocates of both sides and extensive student-centered debate and writing to enhance students' critical, analytical, and communication skills. Topics will include: the feasibility and desirability of a ballistic missile defense system, environmental issues such as the ozone hole and global warming, health issues such as pesticides in food, chemicals in the environment, genetic engineering, and cloning animals.
本项目描述了一个跨学科的五年职业发展计划,涉及物理、化学和生物的研究和教学。在研究中,两种互补的荧光技术将用于研究离子通道(负责神经传导的通道)的构象变化。这些技术,一种测量距离,另一种测量旋转,可以应用于许多其他种类的蛋白质和通道。第一种技术,基于镧系元素的共振能量转移(LRET),是一种已建立的称为荧光共振能量转移技术的改进版本。这是因为镧系元素可以通过一种机制将能量传递给有机染料,这种机制对两个标签之间距离的埃变化很敏感。第二种方法是磷光各向异性的一个版本,利用磷光探针或我们最近发现的偏振发光镧系探针。在该技术中,极化激发脉冲选择性地激发那些与脉冲极化对齐的探针,极化敏感检测监测探针的重新定向。这项技术的优势在于它可以在毫秒的时间尺度上测量旋转运动,这个时间尺度与离子通道中的蛋白质重排有关。LRET将用于测量特定位置放置的镧系原子和有机染料之间的距离,作为细胞膜上电压的函数,并在同时进行电测量时将它们与通道内的运动联系起来。利用磷光各向异性测量适当放置探针的旋转可以进一步表征蛋白质的运动。LRET的初步研究在电压控制离子通道领域取得了里程碑式的进展,首次测量了离子通道蛋白随着电压变化而移动时的实际距离变化。这些结果表明,部分通道是旋转的,这一假设将得到进一步的验证。在教学方面,将开设一门主要面向非科学家的本科课程,题为“科学与公民:当前科学、技术和社会中的争议”。这门课程将以“正反”的形式进行教学,首先是介绍学生对当代科学问题的背景知识,然后是阅读双方支持者的文章,以及广泛的以学生为中心的辩论和写作,以提高学生的批判、分析和沟通技能。议题将包括:弹道导弹防御系统的可行性和可取性,环境问题,如臭氧空洞和全球变暖,健康问题,如食品中的农药,环境中的化学物质,基因工程和克隆动物。
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)
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Paul Selvin其他文献
In vitro and in vivo; kinesin and myosin moving one (or a few) at a time
- DOI:
10.1016/j.bpj.2008.12.1086 - 发表时间:
2009-02-01 - 期刊:
- 影响因子:
- 作者:
Paul Selvin - 通讯作者:
Paul Selvin
High Resolution Imaging Via SHREC And SHRImP For Ultra-High DNA/RNA Resolution
- DOI:
10.1016/j.bpj.2008.12.018 - 发表时间:
2009-02-01 - 期刊:
- 影响因子:
- 作者:
Murat Baday;Ming Xiao;Han Cao;Paul Selvin - 通讯作者:
Paul Selvin
Single Molecule Detection of Transcription Factor using Fluorescent Molecular Beacons
- DOI:
10.1016/j.bpj.2017.11.546 - 发表时间:
2018-02-02 - 期刊:
- 影响因子:
- 作者:
Pin Ren;Yuji Ishitsuka;Paul Selvin - 通讯作者:
Paul Selvin
Measuring the Spatial Arrangement of Nmj-Nachr ion Channel Proteins in the Cell Membrane
- DOI:
10.1016/j.bpj.2010.12.2096 - 发表时间:
2011-02-02 - 期刊:
- 影响因子:
- 作者:
Hannah DeBerg;Nir Friedman;Cong T. Nguyen;Paul Simonson;Paul Selvin - 通讯作者:
Paul Selvin
Advance High Resolution DNA Mapping Technique to Identify Genomic Variations
- DOI:
10.1016/j.bpj.2011.11.2295 - 发表时间:
2012-01-31 - 期刊:
- 影响因子:
- 作者:
Murat Baday;Alex Hastie;Aaron Cravens;Deren E. Kudeki;Ming Xiao;Paul Selvin - 通讯作者:
Paul Selvin
Paul Selvin的其他文献
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{{ truncateString('Paul Selvin', 18)}}的其他基金
EAGER: New Ligand Shells for Small Quantum Dots
EAGER:用于小量子点的新配体壳
- 批准号:
1216342 - 财政年份:2012
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
IDBR: Super-Resolution Made Super-Easy via (Transient-)PhILM
IDBR:通过(瞬态)PhILM 使超分辨率变得超级简单
- 批准号:
1063188 - 财政年份:2011
- 资助金额:
$ 50万 - 项目类别:
Continuing Grant
EAGER: Single Quantum Dots via 2-Photon Excitation
EAGER:通过 2 光子激发的单量子点
- 批准号:
0968976 - 财政年份:2010
- 资助金额:
$ 50万 - 项目类别:
Continuing Grant
IDBR: Instrument Development for In Situ FIONA (Fluorescence Imaging with One Nanometer Accuracy)
IDBR:原位 FIONA(一纳米精度荧光成像)仪器开发
- 批准号:
0649779 - 财政年份:2007
- 资助金额:
$ 50万 - 项目类别:
Continuing Grant
Instrument Development for Imaging and Manipulation of Single Biomolecules
单个生物分子成像和操作的仪器开发
- 批准号:
0215869 - 财政年份:2002
- 资助金额:
$ 50万 - 项目类别:
Standard Grant
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