CFTR-dependent protein interactions regulate diarrhea

CFTR 依赖性蛋白质相互作用调节腹泻

基本信息

批准号：
7614197
负责人：
Anjaparavanda P Naren
金额：
$ 25.7万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-05-15 至 2011-05-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7614197
关键词：
2-Mercaptoethanol Antibodies Basic Science Binding Biological Assay C-terminal Cells Chimeric Proteins Cholera Toxin Complex Coupling Cultured Cells Cyclic AMP-Dependent Protein Kinases Cystic Fibrosis Transmembrane Conductance Regulator Diarrhea Diet Digestive System Disorders Disease Edg4 Protein Epithelial Cells Epithelium Food GTP-Binding Proteins Gastrointestinal tract structure Goals Grant Hand Health Human In Vitro Individual Intestines Knockout Mice Laboratories Lead Length Lysophospholipids Macromolecular Complexes Measurement Mediating Mediator of activation protein Membrane Methods Mission Modeling Molecular Monitor Mus National Institute of Diabetes and Digestive and Kidney Diseases PDZ protein Peptides Physiological Prevention Proteins Recombinant Proteins Recombinants Research Research Personnel Signal Transduction Symptoms Tail Testing Tissues clinically relevant crosslink dithiobis(succinimidylpropionate)improved lysophosphatidic acid mouse model overexpression polypeptide C prevent programs protein protein interaction research study

项目摘要

DESCRIPTION (provided by applicant): The hypothesis to be tested is that lysophosphatidic acid (LPA) inhibits secretory diarrhea through CFTR-dependent protein interactions. The long-term objectives of this laboratory as related to this grant are (i) to gain a better understanding of the dynamic protein-protein interactions that regulate LPA-dependent inhibition of CFTR and (ii) to understand the relevance of these interactions in secretory diarrhea. The specific aims of the grant are (AIM 1) to test the hypothesis that LPA inhibits cholera toxin-induced and CFTR-dependent secretory diarrhea in mice and (AIM 2) to test the hypothesis that a macromolecular complex consisting of LPA2, CFTR, and NHERF2 is required for the LPA-elicited inhibition of CFTR-dependent Cl-transport. To advance the research mission of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), the proposed research will yield important basic science information essential to understanding, treating, and preventing digestive diseases such as secretory diarrhea. In Aim 1, we will test the hypothesis that LPA inhibits cholera toxin-induced and CFTR-dependent secretory diarrhea in mice. In subaim 1a, we will test whether LPA inhibits CFTR function in cultured gut epithelial cells and in excised mouse intestinal tissue. In subaim 1 b, we will test whether LPA inhibits cholera toxin-induced CFTR-dependent secretory diarrhea. In subaim 1c, we will test whether LPA does not inhibit CFTR function in LPA2 receptor knockout mice. In Aim 2, we will test the hypothesis that a macromolecular complex consisting of LPA2, CFTR, and NHERF2 is required for the LPA-elicited inhibition of CFTR-dependent Cl-transport. In subaim 2a, we will determine if LPA2, CFTR, and NHERF2 are assembled in a macromolecular complex in vitro. In subaim 2b, we will cross-link the components of the preexisting macromolecular complex (LPA2, CFTR, and NHERF2) in cultured epithelia and in mouse intestinal epithelial cells. In subaim 2c, we will test whether LPA inhibits the CFTR Cl-transporter due to a physical interaction between LPA2, and CFTR (mediated by NHERF2). At present, the molecular mechanisms responsible for LPA-mediated inhibition of secretory diarrhea are unclear. This project is a critical step in understanding the molecular mechanisms underlying the beneficial effects of LPA, thereby making possible improved treatments in the prevention of secretory diarrhea.

描述（由申请人提供）：要检验的假设是溶血磷脂酸（LPA）通过CFTR依赖性蛋白质相互作用抑制分泌性腹泻。该实验室与该赠款相关的长期目标是（i）更好地理解动态蛋白 - 蛋白质相互作用，这些相互作用调节了LPA依赖性抑制CFTR和（ii），以了解这些相互作用在分泌性腹泻中的相关性。 The specific aims of the grant are (AIM 1) to test the hypothesis that LPA inhibits cholera toxin-induced and CFTR-dependent secretory diarrhea in mice and (AIM 2) to test the hypothesis that a macromolecular complex consisting of LPA2, CFTR, and NHERF2 is required for the LPA-elicited inhibition of CFTR-dependent Cl-transport.为了促进美国国家糖尿病与消化和肾脏疾病（NIDDK）的研究任务，拟议的研究将产生重要的基础科学信息，这对于理解，治疗和预防分泌性腹泻等消化疾病至关重要。在AIM 1中，我们将检验以下假设：LPA抑制小鼠中霍乱毒素诱导的和CFTR依赖性分泌性腹泻。在Subaim 1a中，我们将测试LPA是否抑制培养的肠道上皮细胞和切除的小鼠肠道组织中的CFTR功能。在Subaim 1 B中，我们将测试LPA是否抑制霍乱毒素诱导的CFTR依赖性分泌性腹泻。在Subaim 1C中，我们将测试LPA是否不抑制LPA2受体敲除小鼠中的CFTR功能。在AIM 2中，我们将检验以下假设：LPA引诱抑制CFTR依赖性Cl-Transport需要由LPA2，CFTR和NHERF2组成的大分子复合物。在Subaim 2a中，我们将确定LPA2，CFTR和NHERF2是否在体外组装在大分子复合物中。在Subaim 2b中，我们将在培养的上皮菌和小鼠肠上皮细胞中交叉链接先前存在的大分子复合物（LPA2，CFTR和NHERF2）的成分。在Subaim 2C中，我们将测试LPA是否由于LPA2和CFTR之间的物理相互作用而抑制CFTR CL-Transsterter（由NHERF2介导）。目前，尚不清楚负责LPA介导的分泌性腹泻的分子机制。该项目是理解LPA有益作用的分子机制的关键步骤，从而改善了预防分泌性腹泻的治疗方法。