CAREER: Functional annotation of mitochondrial carrier proteins

1454425Metallo，Christian M.这份职业建议书应用工程方法来生成基本知识，描述线粒体如何在完整的哺乳动物细胞中发挥作用。在地球上几乎所有高等生物中，线粒体既是动力源，又是化学加工厂，提供生命所需的能量和构件。线粒体缺陷会导致多种疾病，包括糖尿病、癌症、神经退行性疾病和衰老。尽管线粒体在能量代谢和生理学中具有重要作用，但线粒体功能的几个方面仍不清楚。例如，我们仍然不知道有多少重要的营养物质和化学物质进入和离开线粒体，在那里它们被分解或合成。使用药物控制这一过程可以治疗多种疾病。这项提案将通过对先进细胞系统进行工程设计和建模来研究线粒体新陈代谢来解决这些问题。这项研究将产生可用于改善人类健康、疾病结果、药物开发和营养的基础知识。代谢生理学研究还将与一项教育计划相结合，以激发学生和教育工作者对生化工程、生物化学和医学的兴趣。为了实现这一目标，建议与当地一所高中共同开发一个以实验室为基础的项目，让学生接触生物工程研究及其在人类生理学中的应用，将酵母代谢与人类疾病(如癌症和糖尿病)联系起来。此外，还将开发一个基于网络的交互工具，以提高更高水平的学生和研究人员学习生物化学和代谢的能力。建议在人类疾病和代谢工程应用的背景下，确定选择性控制真核细胞底物利用的新靶点。线粒体和真核细胞之间的相互作用是地球上最重要的共生关系之一。尽管上个世纪对线粒体结构和代谢活性进行了广泛的研究，但许多线粒体成分的身份和功能仍然是一个谜。稳定同位素示踪和基于系统的代谢通量分析(MFA)为量化低等生物的细胞生理学提供了最有效的手段，但应用于更复杂的物种是复杂的分区。这位研究人员开发了一种新的方法来追踪特定亚细胞间隔中的代谢途径，如线粒体和胞浆。在拟议项目的第一个目标中，这项技术将被用来开发一个全面的、分区的真核代谢和细胞氧化还原潜力模型。细胞工程策略将被应用于从功能上注释将代谢物输送进线粒体和运出线粒体的基因/蛋白质。这些工程细胞最终将使用第一个目标中开发的模型进行表征。这些结果将极大地促进对真核细胞代谢的功能理解，并提高临床医生和工程师控制线粒体功能的能力。这项研究计划将被整合到一个更广泛的教育主题中，旨在教育学生、教师和公众有关生物化学和新陈代谢的知识。这一教育计划包括努力(I)突出STEM领域，特别是工程方法对人类健康、工业和环境的影响；(Ii)与当地一所高中启动基于实验室的计划，使学生接触到生物工程研究及其与人类生理学的相关性；(Iii)开发基于网络的学习工具，作为我们高级设计课程的一部分，以促进和加速学生和研究人员学习代谢途径的相互联系。这些研究和教育活动还将为本科生、研究生和博士后奖学金提供培训机会。这项由CBET部门生物技术和生化工程项目颁发的职业奖项是由分子和细胞生物学部门系统和合成生物学项目共同资助的。

1454425Metallo, Christian M.This CAREER proposal applies engineering approaches to generate fundamental knowledge describing how mitochondria function within intact mammalian cells. Mitochondria serve as both the powerhouse and chemical processing plants in virtually all higher organisms on earth, supplying the energy and building blocks necessary for life. Defects in mitochondria contribute to numerous diseases that include diabetes, cancer, neurodegenerative diseases, and aging. Despite their central importance in energy metabolism and physiology, several aspects of mitochondrial function remain unknown. For example, we still do not know how many important nutrients and chemicals enter and exit the mitochondria where they are broken down or synthesized. Control of such processes using drugs could treat numerous diseases. This proposal will address such questions by engineering and modeling advanced cell systems to study mitochondrial metabolism. This research will generate fundamental knowledge that can be used to improve human health, disease outcomes, drug development, and nutrition. The metabolic physiology research will also be integrated with an educational program that stimulates student and educator interest in biochemical engineering, biochemistry, and medicine. To accomplish this goal development of a lab-based program with a local high school is proposed to expose students to bioengineering research and its application to human physiology, relating yeast metabolism to human diseases such as cancer and diabetes. In addition a web-based interactive tool will be developed to enhance the ability of higher level students and researchers to learn biochemistry and metabolism.It is proposed to identify new targets for selectively controlling substrate utilization in eukaryotes in the context of human disease and metabolic engineering applications. The interplay between mitochondria and eukaryotic cells is one of the most important symbiotic relationships on earth. Despite the extensive studies on mitochondrial structure and metabolic activity conducted over the last century, the identity and function of numerous mitochondrial components remains a mystery. Stable isotope tracing and systems-based metabolic flux analysis (MFA) provide the most effective means of quantifying cellular physiology in lower organisms, but application to more complex species is complicated by compartmentalization. The investigator developed a new approach for tracing metabolic pathways in specific subcellular compartments such as the mitochondria and cytosol. In the first objective within the proposed project this technique will be exploited to develop a comprehensive, compartmentalized model of eukaryotic metabolism and cellular redox potential. Cellular engineering strategies will be applied to functionally annotate the genes/proteins that transport metabolites into and out of mitochondria. These engineered cells will ultimately be characterized using the model developed in the first objective. These results will significantly advance the functional understanding of eukaryotic cell metabolism and improve the ability of clinicians and engineers to control mitochondrial function. This research program will be integrated into a broader educational theme that aims to educate students, teachers, and the public on biochemistry and metabolism. This educational plan includes efforts to (i) highlight the impact of STEM fields and in particular engineering approaches on human health, industry, and the environment; (ii) initiate a lab-based program with a local high school to expose students to bioengineering research and its relevance to human physiology; (iii) develop a web-based learning tool as part of our senior design course to promote and accelerate the learning of metabolic pathway interconnectivity by students and researchers. These research and education activities will also provide training opportunities for undergraduate, graduate, and postdoctoral scholars.This CAREER award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology.