Insulin/TZD regulation of protein structure in fat cells
胰岛素/TZD 对脂肪细胞蛋白质结构的调节
基本信息
- 批准号:6697123
- 负责人:
- 金额:$ 15.15万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2003
- 资助国家:美国
- 起止时间:2003-03-01 至 2005-02-28
- 项目状态:已结题
- 来源:
- 关键词:adipocytesbiological signal transductionconformationdiabetes mellitusfluorescence microscopyfluorescence resonance energy transferfluorescent dye /probegene expressiongreen fluorescent proteinshormone regulation /control mechanismin situ hybridizationinsulinionophoresprotein protein interactionprotein structurethiazolestissue /cell culturetranscription factor
项目摘要
DESCRIPTION (provided by applicant): Diabetes results from deficiencies in insulin production and/or insulin signaling. Insulin signals adipocytes to alter the expression of genes for enzymes and hormones that regulate energy balance. Transcription factors that control the expression of these genes include peroxisome proliferator activated receptor gamma (PPARy) and CCAAT enhancer binding protein alpha (C/EBPa). Insulin signaling alters the phosphorylation status of C/EBPa. PPARy is the receptor for thiazolidenediones (TZDs). PPARy heterodimerizes with RXR, the receptor for 9-cis retinoic acid, which enhances the insulin-sensitizing actions of TZDs. The interactions of these factors with themselves, their ligands and co-activators in response to insulin are poorly defined. Understanding these interactions in the cellular milieu will lead to improved insulin-sensitizing therapies.We uniquely have developed powerful fluorescence microscopy techniques that measure the amounts, structure and interactions of proteins at tens of thousands of locations within cells. Transcription factors and co-factors involved in insulin regulation will be tagged with spectrally distinct derivatives of green fluorescent protein (GFP), and expressed pairwise in 3T3-L1 pre-adipocyte/adipocyte model cells. The relative locations of each factor will be determined microscopically within the living cell by comparing the locations of fluorescence emitted from each fluorophore-factor fusion. Factor location will be compared to gene location by in situ hybridizations in fixed cells. Direct interactions between, and conformations within, the factors will be measured at each subcellular location as the degree to which fluorescence energy excited in the fluorophore attached to one factor (or factor domain) is transferred to a fluorophore attached to the second factor (or domain). Thus, regulation of these cooperative factors will be determined, in 3T3-L1 cells, by measuring the separate and combined effects of insulin, the TZD rosiglitazone and 9-cis retinoic acid on the:1. location, dimerization and interactions of C/EBPa, PPARg, RXRa and their co-activators PGC-1, PGC-2, SRC-la, CBP and TRAP2202. conformation of C/EBPa, PPARg, RXRa and the same co-activators, and3. conformations, dimers and interactions of the above factors specifically in the neighborhood of the genes that they regulate.
描述(由申请人提供):糖尿病是由于胰岛素产生和/或胰岛素信号传递不足所致。胰岛素向脂肪细胞发出信号,改变调节能量平衡的酶和激素基因的表达。控制这些基因表达的转录因子包括过氧化体增殖物激活受体γ(PPARy)和CCAAT增强子结合蛋白α(C/EBPA)。胰岛素信号改变C/EBPA的磷酸化状态。PPARy是噻唑二酮(TZDS)的受体。PPARy与9-顺式维甲酸的受体RXR异源二聚,从而增强TZDS的胰岛素增敏作用。这些因子与自身、它们的配体和共激活剂之间的相互作用对胰岛素的反应尚不清楚。了解细胞环境中的这些相互作用将导致改进的胰岛素增敏疗法。我们独一无二地开发了强大的荧光显微镜技术,可以测量细胞内数万个位置的蛋白质的数量、结构和相互作用。参与胰岛素调节的转录因子和辅助因子将被绿色荧光蛋白(GFP)的不同衍生物标记,并在3T3-L1前脂肪细胞/脂肪细胞模型细胞中成对表达。每个因子的相对位置将通过比较每个荧光团-因子融合发出的荧光位置在活细胞内的显微镜下确定。因子定位将通过固定细胞中的原位杂交与基因定位进行比较。因子之间的直接相互作用和因子内部的构象将在每个亚细胞位置被测量,因为结合在一个因子(或因子结构域)上的荧光团中激发的荧光能量被转移到连接到第二个因子(或因子结构域)上的荧光团的程度。因此,在3T3-L1细胞中,通过测量胰岛素、TZD罗格列酮和9-顺式维甲酸对C/EBPA、PPARg、RXRA及其共激活剂PGC-1、PGC-2、SRC-1a、CBP和TRAP2202的定位、二聚化和相互作用的单独和联合影响,将确定这些协同因子的调节。C/EBPA、PPARg、RXRA和相同的共激活剂的构象;上述因子的构象、二聚体和相互作用,特别是在它们所调控的基因附近。
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)
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Fred J SCHAUFELE的其他文献
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{{ truncateString('Fred J SCHAUFELE', 18)}}的其他基金
Insulin/TZD regulation of protein structure in fat cells
胰岛素/TZD 对脂肪细胞蛋白质结构的调节
- 批准号:
6556578 - 财政年份:2003
- 资助金额:
$ 15.15万 - 项目类别:
Transcription factor interactions at the GH promoter
GH 启动子处的转录因子相互作用
- 批准号:
7185168 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
Transcription factor interactions at the GH promoter
GH 启动子处的转录因子相互作用
- 批准号:
6777165 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
TRANSCRIPTION FACTOR INTERACTIONS AT THE GH PROMOTER
GH 启动子处的转录因子相互作用
- 批准号:
2843564 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
Transcription factor interactions at the GH promoter
GH 启动子处的转录因子相互作用
- 批准号:
6850683 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
TRANSCRIPTION FACTOR INTERACTIONS AT THE GH PROMOTER
GH 启动子处的转录因子相互作用
- 批准号:
6177957 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
TRANSCRIPTION FACTOR INTERACTIONS AT THE GH PROMOTER
GH 启动子处的转录因子相互作用
- 批准号:
6381203 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
Transcription factor interactions at the GH promoter
GH 启动子处的转录因子相互作用
- 批准号:
7367018 - 财政年份:1999
- 资助金额:
$ 15.15万 - 项目类别:
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