STRUCTURE-FUNCTION OF TRANSFORMING GROWTH FACTOR BETA
STRUCTURE-FUNCTION OF TRANSFORMING GROWTH FACTOR BETA
批准号:
2101617
负责人:
LARRY E GENTRY
金额:
$15.05万
依托单位国家:
美国
项目类别:
财政年份:
1993
资助国家:
美国
项目状态:
已结题
起止时间:
1993-08-01 至 1997-05-31
关键词:
affinity chromatography animal tissue bioassay cell differentiation chimeric proteins crosslink enzyme linked immunosorbent assay guinea pigs immunoprecipitation laboratory mouse laboratory rabbit monoclonal antibody polymerase chain reaction protein sequence protein structure function receptor binding receptor expression site directed mutagenesis transforming growth factors western blottings
中文摘要
多功能多肽,转化生长因子-β(TGF-β)
Beta),是从较大的
前体分子。先前的研究表明,亲本的二聚体
一段被称为β-潜伏期相关肽(β-LAP)的片段结合在一个
以非共价方式与较小的二聚体成熟生长因子结合。这
紧密而具体的交互完全归功于测试版,即
在加热和酸处理后变性;热和酸基本上
对成熟生长因子无影响。这样做的总体目标是
建议定义转化生长因子-β分子与
他们的测试圈,并研究潜在的激活机制。
这些研究的结果对于理解
转化生长因子-β生物活性的调节。为了检查每一个测试版-
LAPS,我们将它们在转基因的CHO组织培养细胞中表达
独立于成熟的转化生长因子-β,允许生产大型
生物学的,功能性的测试圈的数量。此绑定源
蛋白质将允许进行测量解离常数的研究
成熟的转化生长因子-β及其多克隆和单抗的生产
抗体。抗体将允许研究评估结构
与其他测试圈的关联性。因为他们基本上不是
针对不同的β-圈反应的抗体,这些免疫球蛋白
将提供新的免疫试剂福特方法,如
免疫沉淀或组织分布研究。此外,
不同的,表达的测试圈将被提纯,使用已经
建立了纯化方案,并用来测试这些络合物是否
结合到细胞表面或细胞外基质上。最后,测试圈
来自转化生长因子-β2a和转化生长因子-β2b,转化生长因子-β3将被表达为
用Beta-Lap嵌合体来研究
延迟。这些研究应该会增进我们对潜伏期的理解
并为转化生长因子-β的调控提供了线索。
英文摘要
The multi-functional polypeptides, transforming growth factor betas (TGF-
betas), are proteolytically derived from the carboxyl-terminus of a larger
precursor molecule. Previous studies indicate that a dimer of the pro
piece, known as beta-latency-associated peptide (beta-LAP), binds in a
noncovalent manner with the smaller, dimeric, mature growth factor. This
tight and specific interaction is entirely due to beta-LAP, being
denatured upon heating and acid treatment; heat and acid have essentially
no effects on the mature growth factor. The overall objective of this
proposal is to define the molecular interaction of the TGF-betas with
their beta-LAPs and to investigate potential activation mechanisms.
Results from these studies are important for the understanding of the
regulation of TGF-beta biological activity. To examine each of the beta-
LAPs, we will express them in transfected CHO tissue culture cells
independent of mature TGF-beta, allowing for the production of large
amounts of a biologically, functional beta-LAP. This source of binding
protein will allow for studies measuring the dissociation constants of
mature TGF-beta and for the production of both polyclonal and monoclonal
antibodies. Antibodies will permit studies to assess the structural
relatedness with other beta-LAPs. Since their are essentially no
antibodies reacting toward the diverse beta-LAPs, these immunoglobulins
will provide new immunereagents ford approaches such as
immuneprecipitation or tissue distribution studies. In addition, the
different, expressed beta-LAPs will be purified, using an already
established purification scheme, and used to test whether these complexes
bind to cell surfaces or to extracellular matrix. Finally, the beta-LAPs
from TGF-beta2a and TGF-beta2b, and TGF-beta3 will be expressed as
chimeras with the beta-LAP to investigate structural regions necessary for
latency. These studies should enhance our understanding of the latent
complex and provide clues to the regulatory controls of the TGF-betas.
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STRUCTURE-FUNCTION OF TRANSFORMING GROWTH FACTOR BETA
-
批准号:3204337
-
项目类别:
-
资助金额:$16.24万
-
财政年份:1993
-
负责人:LARRY E GENTRY
-
依托单位:
STRUCTURE-FUNCTION OF TRANSFORMING GROWTH FACTOR BETA
-
批准号:2101618
-
项目类别:
-
资助金额:$16.04万
-
财政年份:1993
-
负责人:LARRY E GENTRY
-
依托单位:
TRANSFORMING GROWTH FACTOR B1
-
批准号:3459133
-
项目类别:
-
资助金额:$9.28万
-
财政年份:1988
-
负责人:LARRY E GENTRY
-
依托单位:
TRANSFORMING GROWTH FACTOR B1
-
批准号:3459130
-
项目类别:
-
资助金额:$8.95万
-
财政年份:1988
-
负责人:LARRY E GENTRY
-
依托单位:
TRANSFORMING GROWTH FACTOR B1
-
批准号:3459129
-
项目类别:
-
资助金额:$10.41万
-
财政年份:1988
-
负责人:LARRY E GENTRY
-
依托单位:
TRANSFORMING GROWTH FACTOR B1
-
批准号:3459131
-
项目类别:
-
资助金额:$9.14万
-
财政年份:1988
-
负责人:LARRY E GENTRY
-
依托单位:
TRANSFORMING GROWTH FACTOR B1
-
批准号:3459132
-
项目类别:
-
资助金额:$9.29万
-
财政年份:1988
-
负责人:LARRY E GENTRY
-
依托单位:
海外基金