HUMAN LUNG NEUTRAL METALLOENDOPEPTIDASE

申请人建议学习和应用细胞生物学的工具， 分子生物学与人类肺部疾病相关的研究领域。 许多调节肽具有有效的生理促炎作用， 并且生长促进作用局限于肺。中性 金属内肽酶（NEP）似乎在降解许多 这些肽。因此，NEP的调节可能会影响到 肽的效果。 NEP最近被定位于肺， 有证据表明它与反应性气道疾病有关。 长期目标是了解NEP的作用和调节， 正常和患病的人体气道。这项建议的具体目标是 1）从结构上充分表征纯化的人肺NEP， 从功能和免疫学的角度来看。2)从人cDNA中克隆基因 和基因组文库。推导的氨基酸序列可能会提供 关于酶的结构和功能的更多信息。这 这项工作还将提供必要的工具， 转录水平。3)全面评估调制 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方面。第一个将是 上皮细胞潜在调节因子系统筛选 cultures.第二个问题将是审查具体措施的影响。 利用代谢产物合成和降解NEP的调节因素 标记和免疫沉淀。最后的推力将是 将DNA探针纳入研究中，以评估 转录调控 犹他州大学的环境非常适合开发 申请人的调查生涯。在肺科，或 密切配合，是一个有能力的调查小组，具有广泛的， 兴趣从基础生物化学、免疫学和分子生物学， 物理学和临床生理学。肺研究会议，研讨会， 和期刊俱乐部促进了该司的公开交流。提案国 约翰·霍达尔博士和雷·白色博士是这一建议的主要支持者， 调查人员在培养年轻人方面有着出色的记录。 犹他州大学在分子生物学领域特别强大， 生物学和这项提案挖掘了这种潜力。 通过应用分子生物学技术 与其他基础科学教师和学员的互动 具有类似的兴趣，和研究生课程的工作，申请人将获得 分子生物学的坚实基础。

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.