STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
批准号:
6161886
负责人:
R W HARTLEY
金额:
$0.0万
依托单位国家:
美国
项目类别:
财政年份:
--
资助国家:
美国
项目状态:
未结题
起止时间:
至
关键词:
Bacillus bacterial proteins chemical stability enzyme activity enzyme biosynthesis enzyme inhibitors enzyme mechanism enzyme structure esterase inhibitor exoribonucleases gene expression gene mutation genetic regulatory element intermolecular interaction microorganism metabolism molecular cloning nucleic acid sequence protein engineering protein folding protein structure function recombinant DNA
中文摘要
两种蛋白质,芽孢杆菌RNA酶,芽孢杆菌的胞外核糖核酸酶
解淀粉菌及其细胞内抑制剂芽孢杆菌RNA酶抑制剂
作为研究蛋白质折叠和蛋白质-蛋白质的模型系统
交互.芽孢杆菌RNA酶是核糖核酸酶的同源群之一
在原核生物和真核生物中都有发生。重组DNA技术
有三个主要目的:(1)促进生产
野生型和突变型蛋白质;(2)检查结构和
基因的控制序列;和(3)在基因中进行特异性改变。
序列来测试折叠理论和探测barnase-barstar
互动这两种蛋白质现在都可以从重组基因中获得
在大肠在大肠杆菌中,芽孢杆菌RNA酶抑制剂的表达抵消了
芽孢杆菌RNA酶表达这两种蛋白质及其复合物的结构
是已知的。1.5 ℃结构的新晶体学解
埃分辨率提供了新的见解。例如几
可以看到在barnase-barstar界面上的水分子
已经被束缚在自由的barnase上了几对巴纳斯-巴斯塔尔夫妇
在界面中的互补突变,通过体内
选择性技术,已结晶和结构immiment。
Barstar还抑制来自链霉菌菌株的一组RNA酶。这些
酶与arnase的序列同一性仅为
百分之二十五其中,认同度从40%到70%不等。的结构
其中两种,RNA酶Sa和St,是从非重组的工作中已知的。
材料我们现在有了第三种核糖核酸酶Sa 2的结构,
和巴斯塔尔有关系核糖核酸酶基因的重叠片段
St,这是很难抑制barstar,已被克隆,使我们能够
以纠正其公布的氨基酸序列中的几个错误。基因
已克隆并测序了芽孢杆菌RNA酶抑制剂的Sa 2同源物。这
蛋白质抑制所有链霉菌酶,但显然不是
barnase 芽孢杆菌RNA酶基因的可控致死性已被用于
为了有利于产生选择性通用质粒克隆
向量
英文摘要
Two proteins, barnase, the extracellular ribonuclease of Bacillus
amyloliquefaciens, and barstar, its intracellular inhibitor, are used
as a model system for the study of protein folding and protein-protein
interactions. Barnase is one of an homologous group of ribonucleases
ocurring in both prokaryotes and eukaryotes. Recombinant DNA techniques
are being applied with three major aims: (1) to facilitate production
of wild type and mutant proteins; (2) to examine the structural and
contro sequences of the genes; and (3) to make specific changes in the
sequences to test theories of folding and to probe the barnase-barstar
interaction. Both proteins can now be obtained from recombinant genes
in E. coli where expression of barstar counters the lethal effect of
barnase expression. The structures of both proteins and their complex
are known. A new crystallographic solution of the structure at 1.5
angstroms resolution has provided new insights. For example, several
water molecules seen in the barnase-barstar interface can be seen
already bound to free barnase. Several barnase-barstar pairs having
complementary mutations in the interface, obtained by an in vivo
selective technique, have been crystallized and structures are immiment.
Barstar also inhibits a group of RNases from Streptomyces strains. These
enzymes are distantly related to arnase with a sequence identity of only
25%. Among themselves, identify ranges from 40% to 70%. The structures
of two, RNases Sa and St, are known from work on nonrecombinant
material. We now have the structure of a third, RNase Sa2, both alone
and in complex with barstar. Overlapping fragments of the gene for RNase
St, which is poorly inhibited by barstar, have been cloned, allowing us
to correct several errors in its published amino acid sequence. The gene
for Sa2 homolog of barstar has now been cloned and sequenced. This
protein inhibits all the Streptomyces enzymes but apparently not
barnase. The controllable lethality of the barnase gene has been used
to advantage in producing a selective general purpose plasmid cloning
vector
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:5201904
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:3839583
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:3875541
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUENFACIENS
-
批准号:3940232
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUENFACIENS
-
批准号:4689017
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUENFACIENS
-
批准号:3917361
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUENFACIENS
-
批准号:3964011
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:3854534
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:3776059
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:3753948
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
STUDY OF A RIBONUCLEASE AND ITS INHIBITOR FROM BACILLUS AMYLOLIQUEFACIENS
-
批准号:2572771
-
项目类别:
-
资助金额:$0.0万
-
财政年份:--
-
负责人:R W HARTLEY
-
依托单位:
海外基金