13C LABELED GLYCEROL
13C LABELED GLYCEROL
批准号:
6120840
负责人:
DAVID LEMASTER
金额:
$7.79万
依托单位国家:
美国
项目类别:
财政年份:
1999
资助国家:
美国
项目状态:
已结题
起止时间:
1999-01-15 至 2000-01-14
中文摘要
SIR提供D,L-[咪唑-“N_2]组氨酸;LG
甲苯-4-单加氧酶(T4MO)催化NADH和02依赖
甲苯、吲哚、萘、氯仿、
三氯乙烯,一种其他碳氢化合物。该酶复合体包括
由四种成分组成:12.5 kDa Rieske铁氧还蛋白(T4MOC,主要成分
本研究的主题)一个11.6 kDa的调节蛋白(T4MOD);
KDA二铁羟基酶(T4MOH)和一个36 kDa的NADH氧化还原酶。我们
正在用多核高场核磁共振研究T4MOC。我们的计划是
广泛使用为研究其他问题而开发的方法
美国国家磁共振研究所的铁-硫蛋白质
麦迪逊。我们对T4MOC的研究代表了第一个详细的核磁共振
一种Rieske蛋白的研究。因此,它们将提供一个
评估和改进现有核磁共振的实验框架
方法,并将提供有关的电子和动态信息
氧化还原态、质子交换和自旋之间的相关性
可能导致Rieske之间电子转移的离域
和Dfion中心。这将涉及两个截然不同的领域
研究工作:1)超精细位移共振的性质
与T4MOC的氧化和还原状态有关;和2)2-D
和三维核磁共振研究产生的抗磁共振
T4MOC和T4MOD的蛋白质骨架。这项研究的目的是
是为了了解蛋白质(T4MOD)的分子性质,以及
二铁羟基酶(T4MOH),导致100-300倍
催化效率的提高。T4MO系统提供了独特的
研究Rieske之间电子传递的能量学机会
和双铁中心,并研究基于蛋白质的氧合控制
T4MOD等调控蛋白在单个细胞内的催化作用
实验系统。因此,很可能会有重大的新见解
关于二铁加氧酶的热力学和生物控制
将从提议的活动中获得。
英文摘要
The SIR provided D, L-[imidazole-"N2]Histidine; lg
Toluene-4-monooxygenase (T4MO) catalyzes the NADH- and 02-dependent
oxidation of toluenen, indole, naphthalene, chloroform,
trichlorethylene, an dother hydrocarbons. The enzyme complex consists
of four components: a 12.5 kDa Rieske ferredoxin (T4MOC, the primary
subject of this study) an 11.6 kDa regulatory protein (T4MOD); a 216
kDa diiron hydroxylase (T4MOH); and a 36kDa NADH oxidoreductase. We
are studying T4MOC using multinuclear, high field NMR. Our plan is to
make extensive use of methods developed for the study of other
iron-sulfur proteins at the National Magnetic Resonance Facility at
Madison. Our studies of T4MOC represent the first detailed NMR
studies of a Rieske protein. As such, they will provide an
experimental framework for evaluating and improving existing NMR
methods, and will provide eletronic and dynamic information about
correlations between redox states, proton exchange, and spin
delocalization that may contribut to electron transfer between Rieske
and dfiron centers. Two distinct areas will be addressed in this
research effort: 1)properties of hyperfine-shifted resonances
associated with the oxidized and reduced states of T4MOC; and 2) 2-D
and 3-D NMR studies of the diamagnetic resonances arising from the
protein backbones of T4MOC and T4MOD. The objective of this research
is to understand the molecular properties of protein (T4MOD), and the
diiron hydroxylase (T4MOH) that lead to the 100- to 300-fold
enhancements in catalytic rate. The T4MO system offers a unique
opportunity to study the energetics of electron transer between Rieske
and diiron centers, and to study the protein-based control of oxygense
catalysis by regulatory proteins such as T4MOD within a single
experimental system. Thus, it is likely that significan new insights
about thermodynamic and biological control of diiron oxygenase enzymes
will be obtained from the proposed activity.
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13C LABELED GLYCEROL
-
批准号:6309047
-
项目类别:
-
资助金额:$2.74万
-
财政年份:2000
-
负责人:DAVID LEMASTER
-
依托单位:
13C LABELED GLYCEROL
-
批准号:6281464
-
项目类别:
-
资助金额:$2.8万
-
财政年份:1998
-
负责人:DAVID LEMASTER
-
依托单位:
13C LABELED GLYCEROL
-
批准号:6251988
-
项目类别:
-
资助金额:$1.96万
-
财政年份:1997
-
负责人:DAVID LEMASTER
-
依托单位:
海外基金