MICROINJECTION STUDIES ON PROTEIN DEGRADATION
蛋白质降解的微注射研究
基本信息
- 批准号:3274567
- 负责人:
- 金额:$ 21.77万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:1980
- 资助国家:美国
- 起止时间:1980-01-01 至 1994-12-31
- 项目状态:已结题
- 来源:
- 关键词:HeLa cells albumins arginine autophagy biological signal transduction cell bank /registry cell growth regulation cell nucleus cell type chemical conjugate chemical stability conformation cysteine cytoplasm deficient growth media dyes endopeptidases enzyme mechanism enzyme substrate analog globin immunochemistry laboratory mouse laboratory rabbit lysozyme microinjections oxyhemoglobin phenylhydrazines proline protease inhibitor protein folding protein metabolism protein reconstitution protein structure proteolysis radiotracer stress proteins temperature sensitive mutant thermostability tissue /cell culture transport proteins ubiquitin western blottings
项目摘要
The long term objective of this proposal os to elucidate the mechanisms by
which cells selectively degrade their intracellular proteins and to
discover the features of protein that promote their catabolism. Two cell
systems are available in which it is possible to eliminate either proline
endopeptidase or ubiquitin (Ub) conjugation. Individual, radiolabeled
proteins will be injected into each cell line to determine which proteins
are substrates for these pathways. This procedure has already revealed
that ubiquitin and bovine serum albumin ar stabilized in cells lacking
proline endopeptidase and that oxidized hemoglobin is stabilized in ts85
mouse cells which contain a labile Ub-activating enzyme. The half-lives of
a series of temperature-sensitive T4 lysozymes will also be measured to
test the hypothesis that thermodynamic stability correlates with metabolic
stability. Surprisingly, T4 lysozyme is degraded very rapidly in HeLa
cells, and we will determine whether surface cysteines or arginine-arginine
pairs are responsible for its short life. In addition, specific proteins
will be targeted to the nucleus to discover whether their degradation rates
are affected by mislocalization. Finally, ubiquitin will be injected into
ts85 mouse cells or HeLa cells and several aspects of Ub metabolism will be
measured, including Ub's role in autophagy and release of injected
proteins, as well as the subcellular location of Ub conjugates before and
after heat-shock.
这项建议的长期目标是通过以下方式阐明这些机制
哪些细胞选择性地降解它们的细胞内蛋白并
发现促进分解代谢的蛋白质的特性。两个单元格
有一些系统可以消除这两种情况中的任何一种
内肽酶或泛素(Ub)结合。个人,放射性标记
蛋白质将被注射到每个细胞系中,以确定哪些蛋白质
是这些途径的底物。这一过程已经揭示了
泛素和牛血清白蛋白AR在缺乏细胞中稳定
脯氨酸内肽酶和氧化血红蛋白在TS85中稳定
含有一种不稳定的Ub激活酶的小鼠细胞。人类的半衰期
一系列对温度敏感的T4溶菌酶也将被检测到
检验热力学稳定性与新陈代谢相关的假设
稳定性。令人惊讶的是,T4溶菌酶在HeLa中降解得非常快
细胞,我们将确定表面半胱氨酸或精氨酸-精氨酸
Pair对其短暂的寿命负有责任。此外,特定的蛋白质
将针对原子核,以发现它们的降解率
都受到本地化错误的影响。最后,泛素将被注射到
TS85小鼠细胞或HeLa细胞和Ub代谢的几个方面
测量,包括Ub在自噬和注射的释放中的作用
蛋白质,以及Ub结合物的亚细胞定位。
在热震之后。
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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MARTIN C RECHSTEINER其他文献
MARTIN C RECHSTEINER的其他文献
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{{ truncateString('MARTIN C RECHSTEINER', 18)}}的其他基金
Proteasomes, PODs and Polyglutamine Diseases
蛋白酶体、POD 和多聚谷氨酰胺疾病
- 批准号:
6826106 - 财政年份:2004
- 资助金额:
$ 21.77万 - 项目类别:
Proteasomes, PODs and Polyglutamine Diseases
蛋白酶体、POD 和多聚谷氨酰胺疾病
- 批准号:
7056150 - 财政年份:2004
- 资助金额:
$ 21.77万 - 项目类别:
Proteasomes, PODs and Polyglutamine Diseases
蛋白酶体、POD 和多聚谷氨酰胺疾病
- 批准号:
6898835 - 财政年份:2004
- 资助金额:
$ 21.77万 - 项目类别:
Proteasomes, PODs and Polyglutamine Diseases
蛋白酶体、POD 和多聚谷氨酰胺疾病
- 批准号:
7216180 - 财政年份:2004
- 资助金额:
$ 21.77万 - 项目类别:
Proteasomes, PODs and Polyglutamine Diseases
蛋白酶体、POD 和多聚谷氨酰胺疾病
- 批准号:
7391680 - 财政年份:2004
- 资助金额:
$ 21.77万 - 项目类别:
PROPERTIES OF AN ATP-UBIQUITIN-DEPENDENT PROTEASE
ATP 泛素依赖性蛋白酶的特性
- 批准号:
2178633 - 财政年份:1986
- 资助金额:
$ 21.77万 - 项目类别:
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