AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
基本信息
- 批准号:2136612
- 负责人:
- 金额:$ 35.68万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1974
- 资助国家:美国
- 起止时间:1974-09-01 至 1998-08-31
- 项目状态:已结题
- 来源:
- 关键词:active sites aminoacid metabolism carbamoyl phosphate synthetase deficiency endopeptidases enzyme biosynthesis enzyme structure enzyme substrate complex immunochemistry intracellular transport laboratory mouse laboratory rat mitochondria molecular chaperones molecular cloning molecular pathology ornithine carbamoyltransferase orphan disease /drug protein folding protein purification protein transport site directed mutagenesis
项目摘要
The research proposed in this application continues a long-standing
interest in the mechanisms by which nuclearly coded mitochondrial
proteins are targeted, translocated, posttranslocationally
processed, folded, and assembled into their native, active
structures. These processes are central to the maintenance and
propagation of mitochondria, and, hence, to cellular metabolism and
homeostasis. A thorough knowledge and understanding of these
pathways is critical to illuminating the pathogenesis of human
metabolic disease involving mitochondrial enzymes and to the
rational design of strategies for somatic correction of their
deficiencies. The model system used for these studies is intact rat
liver mitochondria and subfractions or components purified
therefrom, and the model substrate is mammalian ornithine
transcarbamylase (OTC), a liver-specific, mitochondrial enzyme
deficient in hyperammonemia. The specific aims include: i)
characterization of the mitochondria processing peptidase (MPP),
one of the enzymes responsible for the posttranslocational
processing of OTC by overexpressing it in E. coli, defining its
active site and metal ion requirements, disrupting its activity by
site-directed mutagenesis, crystallizing it, and solving its
structure; 2) identifying, purifying, and characterizing other
mitochondrial proteases required to cleave a subset of imported
precursors; 3) isolating and cloning leader peptide receptors and
components of the translocation apparatus (contact site) by
recovering translocation complexes, cross-linking translocation
intermediates to members of the apparatus, and purifying identified
proteins immunochemically; 4) exploring the role of individual
amino acid residues in the chaperonin groEL (homologous to the rat
Hsp60 protein) in ATP hydrolysis, substrate protein binding, groES
activation, and protein folding; and 5) determining whether
multimeric enzyme or complex assembly is a protein. mediated
process and what is the nature and identity of the factors required
for this function.
本申请中提出的研究延续了长期以来的
感兴趣的机制,核编码的线粒体
蛋白质被靶向、移位、移位后
经过加工、折叠和组装,
结构.这些过程是维护和
线粒体的繁殖,因此,细胞代谢,
体内平衡深入了解和理解这些
途径对于阐明人类的发病机制至关重要
涉及线粒体酶的代谢疾病,
合理设计的策略,为他们的躯体矫正,
缺陷用于这些研究的模型系统是完整的大鼠
纯化的肝线粒体和亚组分或组分
模型底物是哺乳动物鸟氨酸
转氨甲酰酶(OTC),一种肝脏特异性线粒体酶
缺乏高氨血症。具体目标包括:
线粒体加工肽酶(MPP)的表征,
其中一种酶负责
通过在E.大肠杆菌,定义其
活性位点和金属离子的要求,破坏其活性,
定点突变,使其结晶,并解决其
结构; 2)识别,纯化和表征其他
线粒体蛋白酶所需的切割一个子集的进口
3)分离和克隆前导肽受体,
易位装置的组件(接触部位),
回收易位复合物,交联易位
中间体的装置的成员,并纯化鉴定
蛋白质免疫化学; 4)探索个体的作用
伴侣蛋白groEL中的氨基酸残基(与大鼠同源
Hsp 60蛋白)在ATP水解、底物蛋白结合、groES
激活和蛋白质折叠;以及5)确定是否
多聚体酶或复合物组装体是蛋白质。介导
过程以及所需因素的性质和特性是什么
对于这个功能。
项目成果
期刊论文数量(19)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Import of rat ornithine transcarbamylase precursor into mitochondria: two-step processing of the leader peptide.
- DOI:10.1083/jcb.105.6.2631
- 发表时间:1987-12
- 期刊:
- 影响因子:0
- 作者:Sztul ES;Hendrick JP;Kraus JP;Wall D;Kalousek F;Rosenberg LE
- 通讯作者:Rosenberg LE
Cleavage of precursors by the mitochondrial processing peptidase requires a compatible mature protein or an intermediate octapeptide.
- DOI:10.1083/jcb.113.1.65
- 发表时间:1991-04
- 期刊:
- 影响因子:0
- 作者:Isaya G;Kalousek F;Fenton WA;Rosenberg LE
- 通讯作者:Rosenberg LE
GroEL, GroES, and ATP-dependent folding and spontaneous assembly of ornithine transcarbamylase.
鸟氨酸转氨甲酰酶的 GroEL、GroES 和 ATP 依赖性折叠和自发组装。
- DOI:
- 发表时间:1993
- 期刊:
- 影响因子:0
- 作者:Zheng,X;Rosenberg,LE;Kalousek,F;Fenton,WA
- 通讯作者:Fenton,WA
The general mitochondrial matrix processing protease from rat liver: structural characterization of the catalytic subunit.
大鼠肝脏的一般线粒体基质加工蛋白酶:催化亚基的结构特征。
- DOI:10.1073/pnas.87.20.7978
- 发表时间:1990
- 期刊:
- 影响因子:11.1
- 作者:Kleiber,J;Kalousek,F;Swaroop,M;Rosenberg,LE
- 通讯作者:Rosenberg,LE
Import of the malate dehydrogenase precursor by mitochondria. Cleavage within leader peptide by matrix protease leads to formation of intermediate-sized form.
通过线粒体输入苹果酸脱氢酶前体。
- DOI:
- 发表时间:1988
- 期刊:
- 影响因子:0
- 作者:Sztul,ES;Chu,TW;Strauss,AW;Rosenberg,LE
- 通讯作者:Rosenberg,LE
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FRANTISEK KALOUSEK其他文献
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{{ truncateString('FRANTISEK KALOUSEK', 18)}}的其他基金
AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
- 批准号:
2460746 - 财政年份:1995
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
- 批准号:
2292198 - 财政年份:1995
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
- 批准号:
2292199 - 财政年份:1995
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
- 批准号:
2136611 - 财政年份:1974
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
- 批准号:
3224618 - 财政年份:1974
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM: ENZYME BIOGENESIS AND MUTATION
氨基酸代谢:酶的生物发生和突变
- 批准号:
3224617 - 财政年份:1974
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM--ENZYME BIOGENESIS AND MUTATION
氨基酸代谢——酶的生物发生和突变
- 批准号:
2136610 - 财政年份:1974
- 资助金额:
$ 35.68万 - 项目类别:
AMINO ACID METABOLISM: ENZYME BIOGENESIS AND MUTATION
氨基酸代谢:酶的生物发生和突变
- 批准号:
3224619 - 财政年份:1974
- 资助金额:
$ 35.68万 - 项目类别: