CHARACTERIZATION OF THE COATED VESICLE PROTON PUMP
CHARACTERIZATION OF THE COATED VESICLE PROTON PUMP
批准号:
3285544
负责人:
MICHAEL D FORGAC
金额:
$16.17万
依托单位:
依托单位国家:
美国
项目类别:
财政年份:
1985
资助国家:
美国
项目状态:
已结题
起止时间:
1985-08-30 至 1988-07-31
中文摘要
这项研究的长期目标是确定结构,
依赖三磷酸腺苷的质子泵的分布及其调节机制
包裹着网状蛋白的囊泡。最初的努力将集中在亲和力上
该泵的标识、提纯和改装。
酸化已被确定为加工过程中的一个重要步骤
受体介导的内吞作用后的配体。因此,许多配体
(包括胰岛素、表皮生长因子和去唾液酸糖蛋白)从它们的
低pH值(5.0-6.0)下的受体和受体循环受阻
通过消散pH梯度的试剂。这些药剂还可以阻止感染。
由某些病毒和毒素通过内吞作用进入细胞。这些
结果表明,低pH值激活了配体-受体的解离
受体循环所必需的,而其他研究表明,这是
酸化发生在溶酶体前隔室。
我们已经确定了一种依赖于三磷酸腺苷的质子泵能够酸化
网状蛋白包裹的囊泡,已知的最早的内吞体内隔室
路径。这台泵的部分特点是它的
离子转运特性、周转过程中的磷酸化及抑制物
敏感度。亲和力标签研究将使用
三磷酸腺苷保护抑制剂NBD-C1用于识别三磷酸腺苷结合亚单位
打气筒。洗涤剂增溶蛋白的纯化及重组
将提供有关磷脂小泡的明确信息
该泵的结构、活性及其与其他阳离子的关系
运输ATPase。最后,将产生抗体来对抗纯化的
蛋白质,并用于免疫细胞化学研究以确定
这种泵在细胞内的分布。这些研究对于
我们对这种泵的理解,除了它在
受体介导的内吞作用,可能参与调节
细胞内的pH值和某些病毒和毒素的进入。
英文摘要
The long term goal of this research is to determine the structure,
distribution and mechanism of regulation of the ATP-dependent proton pump
of clathrin-coated vesicles. Initial efforts will be focused on affinity
labeling, purification and reconstitution of this pump.
Acidification has been identified as an essential step in the processing of
ligands following receptor-mediated endocytosis. Thus, a number of ligands
(including insulin, EGF and asialoglycoproteins) dissociate from their
receptors on exposure to low pH (5.0-6.0) and receptor recycling is blocked
by agents which dissipate pH gradients. These agents also block infection
by certain viruses and toxins which enter the cell by endocytosis. These
results suggest that low pH activates the ligand-receptor dissociation
necessary for receptor recycling, and other studies suggest that this
acidification occurs in a prelysosomal compartment.
We have identified an ATP-dependent proton pump capable of acidifying
clathrin-coated vesicles, the earliest known compartment in the endocytic
pathway. This pump has been partially characterized with respect to its
ion transport properties, phosphorylation during turnover and inhibitor
sensitivity. Affinity labeling studies will be carried out using the
ATP-protectable inhibitor NBD-C1 to identify the ATP binding subunit of the
pump. Purification of the detergent solubilized protein and reconstitution
into phospholipid vesicles will provide definitive information on the
structure and activity of this pump and on its relation to other cation
transport ATPases. Finally, antibodies will be raised against the purified
protein and used in immunocytochemical studies to determine the
intracellular distribution of this pump. These studies are essential to
our understanding of this pump which, in addition to its role in
receptor-mediated endocytosis, may be involved in regulation of
intracellular pH and the entry of certain viruses and toxins.
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批准号:10308465
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项目类别:
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资助金额:$18.54万
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财政年份:2020
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负责人:MICHAEL D FORGAC
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依托单位:
Conference--Molecular & Cellular Bioenergetics
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批准号:6597174
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资助金额:$0.6万
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财政年份:2003
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批准号:2177444
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资助金额:$5.7万
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批准号:3304114
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财政年份:1990
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STRUCTURE & PROPERTIES OF THE COATED VESICLE CL CHANNEL
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批准号:3304112
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资助金额:$9.38万
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财政年份:1990
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依托单位:
STRUCTURE & PROPERTIES OF THE COATED VESICLE CL CHANNEL
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批准号:3304113
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项目类别:
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资助金额:$12.9万
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财政年份:1990
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负责人:MICHAEL D FORGAC
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依托单位:
COATED VESICLE PROTON PUMP
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批准号:2177443
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项目类别:
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资助金额:$26.31万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
Structure, Mechanism and Regulation of the V-ATPases
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批准号:6615777
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项目类别:
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资助金额:$46.85万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
CHARACTERIZATION OF THE COATED VESICLE PROTON PUMP
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批准号:3285550
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项目类别:
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资助金额:$16.32万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
CHARACTERIZATION OF THE COATED VESICLE PROTON PUMP
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批准号:3285547
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资助金额:$12.07万
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依托单位:
Structure, Mechanism and Regulation of the V-ATPases
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批准号:7027490
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资助金额:$52.78万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
Structure, mechanism and regulation of the V-ATPases
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批准号:8369970
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项目类别:
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资助金额:$36.3万
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Structure, mechanism and regulation of the V-ATPases
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批准号:8839773
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资助金额:$36.3万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
CHARACTERIZATION OF THE COATED VESICLE PROTON PUMP
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批准号:2177446
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项目类别:
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资助金额:$36.83万
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财政年份:1985
-
负责人:MICHAEL D FORGAC
-
依托单位:
CHARACTERIZATION OF THE COATED VESICLE PROTON PUMP
-
批准号:3285551
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项目类别:
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资助金额:$16.97万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
STRUCTURE, MECHANISM AND REGULATION OF THE V-ATPASES
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批准号:6179660
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项目类别:
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资助金额:$42.73万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
Structure, Mechanism and Regulation of the V-ATPases
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批准号:6525899
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项目类别:
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资助金额:$45.49万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
Structure, Mechanism and Regulation of the V-ATPases
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批准号:6369645
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项目类别:
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资助金额:$46.18万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
STRUCTURE, MECHANISM AND REGULATION OF THE V-ATPASES
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批准号:2749823
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项目类别:
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资助金额:$41.84万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
STRUCTURE, MECHANISM AND REGULATION OF THE V-ATPASES
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批准号:6018630
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项目类别:
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资助金额:$41.49万
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财政年份:1985
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负责人:MICHAEL D FORGAC
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依托单位:
海外基金