CHEMISTRY OF TUBULIN GUANINE NUCLEOTIDE INTERACTIONS
微管蛋白鸟嘌呤核苷酸相互作用的化学性质
基本信息
- 批准号:3289724
- 负责人:
- 金额:$ 11.1万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1985
- 资助国家:美国
- 起止时间:1985-04-01 至 1988-03-31
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
In this application, we outline a series of experiments designid to provide
a more in-depth understanding of the factors controlling the binding and
hydrolytic steps which together constitute the tubulin GTPase reaction and
their relationship to microtubule self-assembly: (1) Examination of the
medium and intermediate oxygen-18 exchange reactions associated with the
assembly-induced GTPase (The goal here is to learn about the reversibility
of certain steps in the hydrolytic mechanism and the rate constants which
define their rates and fluxes.); (2) determination of the stereochemical
course of the hydrolysis using GTP Lambda S labeled with oxygen-16, -17 and
-18 stereospecifically in the Lambda-phosphate (This will help discriminate
between reactions involving a single in-line attack or two inversions, the
latter characteristic of a phosphoryl-protein intermediates); (3)
Characterization of the binding mechanism for guanine nucleotides (The
objective is to learn more about the tubulin conformational changes
preceding or attending ligand binding.); (4) Application of chromium (III)
GTP complexes to define the stereochemistry of tubulin binding of
metal-neucleotide complexes (The approach taken advantage of the invertness
of chromium-nucleotide complexes to ligand exchange to help derive
otherwise nearly inaccessible information about the binding of short-lived
MgGTP2-binding.); (5) Studies of the time-course of hydrolysis during a
single elongation step (This will help to distinguish between prompt and
delayed DTP hydrolysis with repsect to tubulin binding to an elongating
microtubule.); (6) Use of 5'-guanylyl peroxy-diphosphate to potentially
modify the chemical course of GTPase action or to inhibit GTP bining to
tubulin (This would represent the first attempt to utilize another
hydrolyzable analogue of GTP at the exchangeable nucleotide site); (7)
Reexamination of the 32Pi less than greater than GTP and 32Pi less than
greater than GDP exchange reactions of porcine microtubule protein (This
study promises to resolve the conflicting exchange properties of bovine and
porcine brain microtubule proteins); (8) Development of an isothermal
polymerization method to eliminate potentially misleading cold-stable forma
of microtubule protein (This new approach of rapid isothermal concentration
may be helpful in studying the regulation of assembly.). The overall goals
of the project will be realized when results of these and other studies
integrate into a global mechanism.
在本申请中,我们概述了一系列旨在提供
更深入地了解控制约束力的因素,
- 水解步骤,其共同构成微管蛋白GTdR反应,
它们与微管自组装的关系:(1)检测微管的
与氧-18交换反应有关的中等和中等氧-18交换反应
汇编诱导的GTcloud(这里的目标是了解
水解机理中的某些步骤和速率常数
定义它们的速率和通量。)(2)立体化学的测定
使用用氧-16、-17和-18标记的GTP λ S的水解过程
-18立体特异性地在λ-磷酸盐中(这将有助于区分
在涉及一次串联攻击或两次反转的反应之间,
后者是磷酰基蛋白质中间体的特征);(3)
鸟嘌呤核苷酸结合机制的表征(The
目的是了解微管蛋白的构象变化
在配体结合之前或参与配体结合。(4)铬(III)的应用
GTP复合物来定义微管蛋白结合的立体化学,
金属-核苷酸复合物(该方法利用了
铬-核苷酸复合物的配体交换,
否则几乎无法访问的信息绑定的短命
MgGTP 2结合); (5)研究了水解过程中的时间过程,
单伸长步骤(这将有助于区分快速和
延迟的DTP水解与微管蛋白结合到延长的
微管); (6)5 '-鸟苷酰过氧二磷酸的用途,
改变GTP作用的化学过程或抑制GTP比宁,
微管蛋白(这将是首次尝试利用另一种
在可交换的核苷酸位点处的GTP的可水解类似物);(7)
复查32 Pi小于大于GTP和32 Pi小于
大于猪微管蛋白的GDP交换反应(这
这项研究有望解决牛和牛之间相互矛盾的交换特性,
猪脑微管蛋白);(8)等温
消除潜在误导的冷稳定形式的聚合方法
微管蛋白的快速等温浓缩新方法
可能有助于研究组装的调节)。 总目标
当这些研究和其他研究的结果出来时,
纳入全球机制。
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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Daniel Lee Purich其他文献
Daniel Lee Purich的其他文献
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{{ truncateString('Daniel Lee Purich', 18)}}的其他基金
ACTIN-BASED MOTILITY BY CLAMPED-FILAMENT MOTORS
钳位灯丝电机基于肌动蛋白的运动
- 批准号:
6731384 - 财政年份:2004
- 资助金额:
$ 11.1万 - 项目类别:
ACTIVE-BASED MOTILITY BY CLAMPED-FILAMENT MOTORS
通过夹紧灯丝电机实现主动运动
- 批准号:
6879064 - 财政年份:2004
- 资助金额:
$ 11.1万 - 项目类别:
ACTIVE-BASED MOTILITY BY CLAMPED-FILAMENT MOTORS
通过夹紧灯丝电机实现主动运动
- 批准号:
7039166 - 财政年份:2004
- 资助金额:
$ 11.1万 - 项目类别:
TUBULIN BOUND DEOXY GTP AND NGF TREATED NEURONS
微管蛋白结合脱氧 GTP 和 NGF 处理的神经元
- 批准号:
2431223 - 财政年份:1995
- 资助金额:
$ 11.1万 - 项目类别:
TUBULIN BOUND DEOXY GTP AND NGF TREATED NEURONS
微管蛋白结合脱氧 GTP 和 NGF 处理的神经元
- 批准号:
2270715 - 财政年份:1995
- 资助金额:
$ 11.1万 - 项目类别:
TUBULIN BOUND DEOXY GTP AND NGF TREATED NEURONS
微管蛋白结合脱氧 GTP 和 NGF 处理的神经元
- 批准号:
2270716 - 财政年份:1995
- 资助金额:
$ 11.1万 - 项目类别:
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