Kinase Regulation of the Epithelial Sodium Channel

上皮钠通道的激酶调节

基本信息

批准号：
7209011
负责人：
JONATHAN J LEBOWITZ
金额：
$ 12.65万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-05-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7209011
关键词：
A549 Affect Agonist Aldosterone Alveolar Cell Amiloride Apical Bacteria Basic Science Binding Biochemical Biological Assay Carcinoma Cell Adhesion Molecules Cell Line Cell Nucleus Cell physiology Cell surface Cells Chimeric Proteins Cleaved cell Clinical Condition DNA DNA Binding Domain Data Dexamethasone Dominant-Negative Mutation Down-Regulation Duct (organ) structure Electrodes Environment Epithelial Epithelium Event Faculty Feedback Genes Genetic Transcription Glucocorticoid Receptor Glucocorticoids Glutathione S-Transferase Goals Grant Heat-Shock Response Human I Kappa B-Alpha Immunoprecipitation In Vitro Interleukin-1 Interleukin-1 Receptors Kidney Label Laboratories Ligands Link Lung Measures Mediating Mentors Messenger RNA Methionine Methods Mineralocorticoids Modeling Molecular Monitor Mus Mutate NF-kappa B Na(+)-K(+)-Exchanging ATPase Nature Nephrology Oocytes Oxidative Stress Pathway interactions Phosphorylation Phosphorylation Inhibition Phosphotransferases Physiological Positioning Attribute Prevention Probability Promoter Regions Protein Overexpression Proteins Receptor Activation Regulation Relative (related person)Reporter Reporter Genes Research Research Personnel Research Project Grants Role Serine Signal Pathway Signal Transduction Site Sodium Channel Steroids Stimulus Surface System Technical Expertise Techniques Threonine Tissues Translating Tumor Necrosis Factor Receptor Tumor Necrosis Factor-Beta Viral Voltage-Clamp Technics Western Blotting Xenopus oocyte alveolar type II cell base career chemokine epithelial Na+ channel inhibitor/antagonist interest multicatalytic endopeptidase complex mutant novel protein protein interaction receptor research study skills vector vector control yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Dr. Lebowitz is currently conducting research in the laboratory of John P. Johnson at the Univeristy of Pittsburgh. His career goals for the period of this grant are to further develop both the technical expertise (e.g. in molecular, biochemical, and electrophysiological methods) and in the creative skills necessary to become a successful independent investigator. After 1-2 years of mentored research he plans to make the transition to a tenure-track faculty position within the renal division. His long term goal is to become a fully independent academic investigator performing basic research in an environment where bench results may be applied to important clinical problems within the broad fields of nephrology and cellular physiology. He is especially interested in the regulation of epithelial transport systems. The basis for this research project is a the discovery of a novel interaction between the beta subunit of the kinase that cleaves the inhibitor of nuclear factor kappa B (IKKbeta) and the beta subunit of the epithelial sodium channel (ENaC). Using the yeast two hybrid system and immunoprecipitation techniques, we have established this interaction. Further, using the two electrode voltage clamping technique and co-expression of IKKbeta and all three ENaC subunits in Xenopus oocytes, we have shown that this interaction increases ENaC activity. Although studies have linked activation of nuclear factor kappa B (NF-kappaB) with changes in ENaC activity, no study has elucidated the nature of this relationship nor has any research explored the basic mechanism involved. Since our results show that ENaC may be activated through the NF-kappaB cascade as well as regulated by glucocorticoids and mineralocorticoids, an interaction between these two pathways is apparent. Preliminary data demonstrates that mineralocorticoids may also activate pathways in renal and pulmonary epithelia.

描述（由申请人提供）： Lebowitz 博士目前正在匹兹堡大学 John P. Johnson 实验室进行研究。他在这笔资助期间的职业目标是进一步发展技术专长（例如分子、生物化学和电生理学方法）和成为一名成功的独立研究者所需的创造性技能。经过 1-2 年的指导研究后，他计划过渡到肾脏科的终身教职。他的长期目标是成为一名完全独立的学术研究者，在将实验室结果应用于肾病学和细胞生理学广泛领域内的重要临床问题的环境中进行基础研究。他对上皮运输系统的调节特别感兴趣。该研究项目的基础是发现裂解核因子 kappa B 抑制剂 (IKKbeta) 的激酶 β 亚基与上皮钠通道 (ENaC) β 亚基之间的新型相互作用。使用酵母两种杂交系统和免疫沉淀技术，我们已经建立了这种相互作用。此外，使用两电极电压钳技术以及 IKKbeta 和非洲爪蟾卵母细胞中所有三个 ENaC 亚基的共表达，我们发现这种相互作用会增加 ENaC 活性。尽管研究已将核因子 kappa B (NF-kappaB) 的激活与 ENaC 活性的变化联系起来，但没有研究阐明这种关系的本质，也没有任何研究探索所涉及的基本机制。由于我们的结果表明 ENaC 可能通过 NF-kappaB 级联被激活，并受到糖皮质激素和盐皮质激素的调节，因此这两种途径之间的相互作用是显而易见的。初步数据表明，盐皮质激素也可能激活肾和肺上皮细胞的通路。