HUMAN LUNG NEUTRAL METALLOENDOPEPTIDASE

申请人建议学习和应用细胞生物学的工具和 分子生物学是与人类肺部疾病相关的研究领域。 一些具有强大的生理性促炎作用的调节肽， 而促进生长的作用已经局限于肺。中立者 金属内肽酶(NEP)似乎在降解许多 这些多肽。因此，NEP的调制可能会影响到 多肽的作用。NEP最近被定位于肺和 有证据表明，它与反应性呼吸道疾病有关。 长期目标是了解NEP在体内的作用和调节 正常的和患病的人类呼吸道。这项提议的具体目的是 目的：1)从结构上充分鉴定纯化的人肺NEP， 从功能和免疫学角度来看。2)克隆人c DNA基因 和基因组文库。推导出的氨基酸序列可能会提供 关于酶的结构和功能的其他信息。这 这项工作还将提供必要的工具，以审查对NEP的监管 转录水平。3)综合评估调制方式 上皮细胞NEP从转录水平到翻译后水平。 这项工作将包括三个主要推动力。第一个将是一个 系统筛选上皮细胞潜在调控因子 文化。第二个将是对具体影响的审查 利用代谢途径调节NEP合成和降解的因素 标记和免疫沉淀。最后的推力将是一个 将DNA探针纳入研究以评估 转录调控。 犹他大学的环境非常适合开发 申请人的调查职业。在肺科，或 紧密结合，是一支有能力的调查小组，拥有广泛的 从基础生物化学、免疫学和分子生物学到 物理学和临床生理学。肺部研究会议、研讨会、 杂志俱乐部促进了该部门的公开交流。赞助商 约翰·霍达尔博士和雷·怀特博士对这项建议非常有成效 在培养年轻人方面有良好记录的调查人员。 犹他大学在分子研究领域尤其强大 生物学和这项提议挖掘了这一潜力。 通过将分子生物学技术应用于 建议，与其他基础科学教员和实习生的互动 有相似的兴趣和研究生课程，申请者将获得 扎实的分子生物学基础。

The applicant proposes to learn and apply the tools of cell biology and molecular biology to a research area relevant to human lung disease. A number of regulatory peptides with potent physiologic pro-inflammatory, and growth promoting effects have been localized to the lung. A neutral metalloendopeptidase (NEP) appears to play a key role in degrading many of these peptides. The modulation of NEP will thus likely impact on the extent of the peptides' effects. NEP has recently been localized to the lung and evidence suggests that it is relevant to reactive airways disease. The long term objective is to understand the role and regulation of NEP in normal and diseased human airways. The specific aims of this proposal are to: 1) Fully characterize the purified human lung NEP from a structural, functional, and immunological standpoint. 2) Clone the gene from human cDNA and genomic libraries. The deduced amino acid sequence will likely provide additional information on the structure and function of the enzyme. This work will also provide the tools necessary to examine regulation of NEP at the transcriptional level. 3) Comprehensively assess modulation of epithelial cell NEP from the transcriptional to post-translational level. This work will consist of three major thrusts. The first will be a systematic screening of potential modulating factors in epithelial cell cultures. The second will be an examination of the effects of specific modulating factors on synthesis and degradation of NEP utilizing metabolic labelling and immunoprecipitation. The final thrust will be an incorporation of the DNA probe into the studies for an assessment of transcriptional regulation. The environment at the University of Utah is ideal for developing the investigative career of the applicant. Within the Pulmonary Division, or closely aligned, are a capable group of investigators with a wide range of interests from basic biochemistry, immunology, and molecular biology to physics and clinical physiology. Pulmonary research conferences, seminars, and journal clubs foster open communication in the Division. The sponsors of this proposal, Dr. John Hoidal and Dr. Ray White, are highly productive investigators with an excellent track record in developing young people. The University of Utah is particularly strong in the area of molecular biology and this proposal taps that potential. Through the application of the techniques of molecular biology to this proposal, the interaction with other basic science faculty and trainees with similar interests, and graduate course work, the applicant will gain a solid foundation in molecular biology.