FOLDING AND ASSEMLY IN VISUAL RHODOPSIN
FOLDING AND ASSEMLY IN VISUAL RHODOPSIN
批准号:
6179332
负责人:
KEVIN Donald RIDGE
金额:
$10.36万
依托单位国家:
美国
项目类别:
财政年份:
1996
资助国家:
美国
项目状态:
已结题
起止时间:
1996-06-01 至 2002-05-31
关键词:
Escherichia coli SDS polyacrylamide gel electrophoresis analytical ultracentrifugation calorimetry circular dichroism cow fluorescence spectrometry gene complementation genetic manipulation protein biosynthesis protein folding protein reconstitution receptor expression rhodopsin rod cell site directed mutagenesis thermodynamics ultraviolet spectrometry visual photoreceptor western blottings
中文摘要
描述(改编自申请人的摘要):蛋白质的研究
折叠过程提供了对机制、动力学和
多肽折叠的热力学。 这项研究的目标是
提供杆状电池折叠和组装的详细分析
光感受器视紫红质通过专注于识别和
牛视蛋白独立折叠域的表征
脱辅基蛋白。 通过基因操作产生的视蛋白多肽片段
已对牛视蛋白基因的体内功能组装进行了检查。
值得注意的是,两个或三个互补视蛋白多肽的共表达
在细胞质区域分离的片段允许形成
视紫红质具有与天然色素相似的光谱特征。
这些结果表明视紫红质的功能组装可能是
由多个蛋白质折叠结构域的联合介导。 为了进一步
证实这些发现,其他网站的本地化
视蛋白多肽链的不连续性允许体内互补
正在被追捕。 体外补充系统的开发将
通过检查促进视蛋白多肽的条件来促进
片段复合物的形成并通过其结构表征
圆二色性 (CD) 和荧光光谱的特性
通过分析超速离心作为它们的结合状态。 按顺序
产生足够量的视蛋白多肽片段用于进一步
互补研究正在进行中,细菌过度表达。 的
伴随片段互补的相互作用的性质和程度
自然和定点突变对该过程的影响将
通过滴定量热法进行检查。 分析正在发生的转变
通过差示扫描量热法观察视蛋白多肽片段
应验证它们是否由一个或多个独立折叠组成
域。 除了识别含有牛视蛋白的区域外
足够的信息来独立折叠、插入膜中,以及
组装成功能分子,这些研究应该提供见解
与一些自然发生的结构后果相关
在患有色素性视网膜炎的患者中观察到视蛋白突变。 他们
预计也与不断增长的折叠和组装有关
与鸟嘌呤核苷酸结合偶联的相关受体的数量
蛋白质。
英文摘要
DESCRIPTION (Adapted from applicant's abstract): Studies of the protein
folding process offer insights into the mechanisms, kinetics, and
thermodynamics of polypeptide folding. The goal of this research is to
provide a detailed analysis of the folding and assembly of the rod cell
photoreceptor rhodopsin by focusing on the identification and
characterization of independent folding domains in the bovine opsin
apoprotein. Opsin polypeptide fragments generated by genetic manipulation
of the bovine opsin gene have been examined for functional assembly in vivo.
Remarkably, co-expression of two or three complementary opsin polypeptide
fragments separated in the cytoplasmic region allows the formation of
rhodopsins with spectral characteristics similar to the native pigment.
These results suggest that the functional assembly of rhodopsin may be
mediated by the association of multiple protein-folding domains. To further
substantiate these findings, the localization of additional sites where
discontinuity of the opsin polypeptide chain allows in vivo complementation
is being pursued. The development of an in vitro complementing system will
be facilitated by examining conditions which promote opsin polypeptide
fragment complex formation and through characterization of their structural
properties by circular dichroism (CD) and fluorescence spectroscopy as well
as their state of association by analytical ultracentrifugation. In order
to produce sufficient quantities of opsin polypeptide fragments for further
complementation studies, bacterial overexpression is in progress. The
nature and extent of the interactions accompanying fragment complementation
and the effects of natural and site-directed mutations on the process will
be examined by titration calorimetry. Analysis of the unfolding transitions
in the opsin polypeptide fragments by differential scanning calorimetry
should verify whether they are composed of one or more independent folding
domains. In addition to identifying regions of bovine opsin which contain
sufficient information to independently fold, insert into a membrane, and
assemble into a functional molecule, these studies should provide insights
into the structural consequences associated with some naturally-occurring
opsin mutations seen in patients afflicted with retinitis pigmentosa. They
are also expected to have relevance to the folding and assembly of a growing
number of related receptors which are coupled to guanine nucleotide-binding
proteins.
期刊论文(8)
专著(0)
科研奖励(0)
会议论文
Heterologous expression of bovine opsin in Pichia pastoris.
牛视蛋白在毕赤酵母中的异源表达。
DOI:
10.1016/s0076-6879(00)15831-8
发表时间:
2000
期刊:
Methods in enzymology
影响因子:
--
作者:
[Abdulaev,NG, Ridge,KD]
通讯作者:
Ridge,KD
DOI:
10.1074/jbc.274.30.21437
发表时间:
1999
期刊:
The Journal of biological chemistry
影响因子:
--
作者:
[Ridge,KD, Ngo,T, Lee,SS, Abdulaev,NG]
通讯作者:
Abdulaev,NG
Folding and assembly of rhodopsin from expressed fragments.
来自表达片段的视紫红质的折叠和组装。
DOI:
10.1016/s0076-6879(00)15834-3
发表时间:
2000
期刊:
Methods in enzymology
影响因子:
--
作者:
[Ridge,KD, Abdulaev,NG]
通讯作者:
Abdulaev,NG
Structural Analysis of the Rhodopsin-Transducin Complex
-
批准号:7248586
-
项目类别:
-
资助金额:$33.49万
-
财政年份:2006
-
负责人:KEVIN Donald RIDGE
-
依托单位:
Structural Analysis of the Rhodopsin-Transducin Complex
-
批准号:7096311
-
项目类别:
-
资助金额:$35.71万
-
财政年份:2006
-
负责人:KEVIN Donald RIDGE
-
依托单位:
Structural Analysis of the Rhodopsin-Transducin Complex
-
批准号:7409551
-
项目类别:
-
资助金额:$32.82万
-
财政年份:2006
-
负责人:KEVIN Donald RIDGE
-
依托单位:
STRUCTURAL STUDIES OF G-PROTEIN COUPLED RECEPTORS
-
批准号:6708869
-
项目类别:
-
资助金额:$1.76万
-
财政年份:2001
-
负责人:KEVIN Donald RIDGE
-
依托单位:
STRUCTURAL STUDIES OF G-PROTEIN COUPLED RECEPTORS
-
批准号:6628607
-
项目类别:
-
资助金额:$22.2万
-
财政年份:2001
-
负责人:KEVIN Donald RIDGE
-
依托单位:
STRUCTURAL STUDIES OF G-PROTEIN COUPLED RECEPTORS
-
批准号:6498231
-
项目类别:
-
资助金额:$22.2万
-
财政年份:2001
-
负责人:KEVIN Donald RIDGE
-
依托单位:
STRUCTURAL STUDIES OF G-PROTEIN COUPLED RECEPTORS
-
批准号:6879356
-
项目类别:
-
资助金额:$16.32万
-
财政年份:2001
-
负责人:KEVIN Donald RIDGE
-
依托单位:
STRUCTURAL STUDIES OF G-PROTEIN COUPLED RECEPTORS
-
批准号:6233046
-
项目类别:
-
资助金额:$22.2万
-
财政年份:2001
-
负责人:KEVIN Donald RIDGE
-
依托单位:
FOLDING AND ASSEMLY IN VISUAL RHODOPSIN
-
批准号:2711140
-
项目类别:
-
资助金额:$10.36万
-
财政年份:1996
-
负责人:KEVIN Donald RIDGE
-
依托单位:
FOLDING AND ASSEMLY IN VISUAL RHODOPSIN
-
批准号:2165388
-
项目类别:
-
资助金额:$4.94万
-
财政年份:1996
-
负责人:KEVIN Donald RIDGE
-
依托单位:
FOLDING AND ASSEMLY IN VISUAL RHODOPSIN
-
批准号:2888476
-
项目类别:
-
资助金额:$10.36万
-
财政年份:1996
-
负责人:KEVIN Donald RIDGE
-
依托单位:
FOLDING AND ASSEMLY IN VISUAL RHODOPSIN
-
批准号:2430391
-
项目类别:
-
资助金额:$10.26万
-
财政年份:1996
-
负责人:KEVIN Donald RIDGE
-
依托单位:
FOLDING AND ASSEMBLY IN VISUAL RHODOPSIN
-
批准号:2165387
-
项目类别:
-
资助金额:$10.0万
-
财政年份:1995
-
负责人:KEVIN Donald RIDGE
-
依托单位:
MOLECULAR BASIS FOR TRANSMEMBRANE SIGNALING IN RHODOPSIN
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批准号:3039386
-
项目类别:
-
资助金额:$2.86万
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财政年份:1992
-
负责人:KEVIN Donald RIDGE
-
依托单位:
MOLECULAR BASIS FOR TRANSMEMBRANE SIGNALING IN RHODOPSIN
-
批准号:3039385
-
项目类别:
-
资助金额:$2.27万
-
财政年份:1991
-
负责人:KEVIN Donald RIDGE
-
依托单位:
MOLECULAR BASIS FOR TRANSMEMBRANE SIGNALING IN RHODOPSIN
-
批准号:3039384
-
项目类别:
-
资助金额:$2.0万
-
财政年份:1990
-
负责人:KEVIN Donald RIDGE
-
依托单位: