G PROTEINS AND PAROTID SECRETORY GRANULE
G PROTEINS AND PAROTID SECRETORY GRANULE
批准号:
2865241
负责人:
EILEEN L WATSON
金额:
$26.24万
依托单位:
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:
1993-09-01 至 2003-08-31
关键词:
ADP ribosylation G protein acinar cell amylases cell membrane chloride channels cytoplasm electrical conductance electron microscopy exocrine glands exocytosis granule immunoelectron microscopy immunofluorescence technique laboratory rat light microscopy membrane permeability membrane transport proteins parotid gland phosphorylation protein structure function radiotracer secretion tissue /cell culture western blottings
中文摘要
拟议研究的总体目标是了解
唾液外分泌细胞中GTP结合蛋白的功能/作用
从我们实验室获得的证据表明,异三聚体
单体G蛋白定位于大鼠腮腺腺泡细胞
分泌颗粒和质膜上也是如此。假设是
在这个方案中测试的是位于分泌颗粒上的G蛋白
膜的功能是调节离子通道的门控和胞吐。
在大鼠腮腺中。具体目标是:
L)异三聚体和单体G的鉴定和定位
蛋白质。G蛋白将在不同的细胞组分中被鉴定
包括质膜、胞浆和分泌颗粒膜。
[32P]ADP-核糖化双向凝胶电泳法,
放射性标记GTP与特异性G蛋白的结合及免疫印迹分析
抗血清。G蛋白在腮腺组织切片和腮腺组织中的定位
腺泡细胞将通过免疫组织化学/免疫细胞化学(LIGH
和电子显微镜)。
2)确定G蛋白对氯离子电导的作用
分离的分泌颗粒。对于这些研究,GTP、GTP类似物、A1F4、
细菌毒素,ADP-核糖化因子(ARF),特异性G蛋白
抗血清和影响单体G蛋白释放/激活的试剂
将会被利用。G蛋白对氯离子电导的调节作用
磷酸化/去磷酸化机制也将在
用放射性标记技术分离颗粒和通透性细胞
[32P]ATP和[32P]正磷酸盐。
3)检测G蛋白在调节分泌中的作用
(胞吐)来自腮腺腺泡细胞。GTP和GTP类似物的作用
在分泌的后期,即淀粉酶的释放将在
通透性细胞。分泌物中G蛋白的易位
其他细胞隔间的颗粒将在腺泡细胞中确定。
在细胞刺激后;移位将通过
免疫荧光/免疫电子显微镜、免疫印迹分析和
放射性标记的GTP结合。所获得的结果将与淀粉酶相关联
放手。
拟议的研究应提供有关
G蛋白的特性以及对其功能的重大洞察
在唾液外分泌细胞中。这些研究还将提供一个框架
用于检查与以下疾病有关的细胞生化缺陷
唾液功能障碍。
英文摘要
The overall goal of the proposed research is to understand the
function/role of GTP-binding proteins in salivary exocrine cells.
Evidence obtained from Our laboratory suggests that both heterotrimeric
and monomeric G proteins are located on the rat parotid acinar cell
secretory granule as well as on the plasma membranes. The hypothesis to be
tested in this proposal is that G proteins, located on secretory granule
membranes function to modulate the gating of ion channels and exocytosis
in the rat parotid gland. Specific aims are:
l) Identification and localization of heterotrimeric and monomeric G
proteins. G proteins will be identified in various cellular fractions
including the plasma membrane, cytosol and secretory granule membranes by
two-dimensional gel electrophoresis using [32P] ADP-ribosylation,
radiolabeled GTP binding and immunoblot analysis with specific G protein
antisera. Localization of G proteins in parotid tissue sections and in
acinar cells will be by immunohistochemistry/immunocytochemistry (light
and electron microscopy).
2) Determination of a role for G proteins on the Cl- conductance of
isolated secretory granules. For these studies GTP, GTP analogs, A1F4,
bacterial toxins, ADP-ribosylating factor (ARF), specific G protein
antisera, and agents that affect monomeric G protein release/activation
will be utilized. Modulation of G protein-regulated Cl- conductance by
phosphorylation/dephosphorylation mechanisms will also be examined in
isolated granules and in permeabilized cells by using radiolabeled
[32P]ATP and [32P]orthophosphate, respectively.
3) Examination of the role of G proteins in regulating secretion
(exocytosis) from parotid acinar cells. The effects of GTP and GTP analogs
on the late stage of secretion, i.e. amylase release will be determined in
permeabilized cells. The translocation of G proteins from secretory
granules to other cellular compartments will be determined in acinar cells
following cell stimulation; translocation will be examined by
immunofluorescence/immunoelectron microscopy, immunoblot analysis and
radiolabeled GTP binding. Results obtained will be correlated with amylase
release.
The proposed research should provide significant information on the
identity of G proteins as well as significant insight as to their function
in salivary exocrine cells. These studies will also provide a framework
for examining cellular biochemical defects in diseases relating to
salivary dysfunction.
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会议论文
G PROTEINS AND PAROTID SECRETORY GRANULE
-
批准号:6523830
-
项目类别:
-
资助金额:$29.69万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND RAT PAROTID SECRETORY GRANULE
-
批准号:2131643
-
项目类别:
-
资助金额:$23.01万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND PAROTID SECRETORY GRANULE
-
批准号:2897045
-
项目类别:
-
资助金额:$27.87万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND RAT PROTID SECRETORY GRANULE
-
批准号:3567992
-
项目类别:
-
资助金额:$21.98万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND RAT PAROTID SECRETORY GRANULE
-
批准号:2518118
-
项目类别:
-
资助金额:$24.84万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND RAT PAROTID SECRETORY GRANULE
-
批准号:3482846
-
项目类别:
-
资助金额:$21.98万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND PAROTID SECRETORY GRANULE
-
批准号:6176170
-
项目类别:
-
资助金额:$27.99万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND RAT PAROTID SECRETORY GRANULE
-
批准号:2131644
-
项目类别:
-
资助金额:$23.89万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND PAROTID SECRETORY GRANULE
-
批准号:6379756
-
项目类别:
-
资助金额:$28.83万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
G PROTEINS AND RAT PAROTID SECRETORY GRANULE
-
批准号:2131642
-
项目类别:
-
资助金额:$25.62万
-
财政年份:1993
-
负责人:EILEEN L WATSON
-
依托单位:
DEVELOPMENT OF DUCTAL CELL CULTURE FROM PAROTID GLAND
-
批准号:2131423
-
项目类别:
-
资助金额:$3.76万
-
财政年份:1992
-
负责人:EILEEN L WATSON
-
依托单位:
DEVELOPMENT OF DUCTAL CELL CULTURE FROM PAROTID GLAND
-
批准号:3425863
-
项目类别:
-
资助金额:$3.7万
-
财政年份:1992
-
负责人:EILEEN L WATSON
-
依托单位:
SUGAR METABOLISM IN ORAL ACTINOMYCES VISCOSUS
-
批准号:3222085
-
项目类别:
-
资助金额:$16.97万
-
财政年份:1988
-
负责人:EILEEN L WATSON
-
依托单位:
SUGAR METABOLISM IN ORAL ACTINOMYCES VISCOSUS
-
批准号:3222088
-
项目类别:
-
资助金额:$16.72万
-
财政年份:1988
-
负责人:EILEEN L WATSON
-
依托单位:
SUGAR METABOLISM IN ORAL ACTINOMYCES VISCOSUS
-
批准号:3222086
-
项目类别:
-
资助金额:$15.91万
-
财政年份:1988
-
负责人:EILEEN L WATSON
-
依托单位:
SUGAR METABOLISM IN ORAL ACTINOMYCES VISCOSUS
-
批准号:3222087
-
项目类别:
-
资助金额:$16.19万
-
财政年份:1988
-
负责人:EILEEN L WATSON
-
依托单位:
SALIVARY SECRETION--ROLE OF CALCIUM
-
批准号:2129061
-
项目类别:
-
资助金额:$25.76万
-
财政年份:1979
-
负责人:EILEEN L WATSON
-
依托单位:
SALIVARY SECRETION--ROLE OF CALCIUM
-
批准号:3219301
-
项目类别:
-
资助金额:$17.05万
-
财政年份:1979
-
负责人:EILEEN L WATSON
-
依托单位:
SALIVARY SECRETION-ROLE OF CALCIUM
-
批准号:3219304
-
项目类别:
-
资助金额:$25.03万
-
财政年份:1979
-
负责人:EILEEN L WATSON
-
依托单位:
Salivary secretion-role of calcium
-
批准号:6727425
-
项目类别:
-
资助金额:$29.41万
-
财政年份:1979
-
负责人:EILEEN L WATSON
-
依托单位:
海外基金