CAREER: Functional annotation of mitochondrial carrier proteins

[1454425] m . metallo, Christian m .这个职业建议应用工程方法来生成描述完整哺乳动物细胞内线粒体功能的基础知识。在地球上几乎所有的高等生物中，线粒体既是发电站，又是化学加工厂，为生命提供必要的能量和基础材料。线粒体缺陷会导致许多疾病，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上具有重要意义，但线粒体功能的几个方面仍不为人所知。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出线粒体，在那里它们被分解或合成。用药物控制这些过程可以治疗许多疾病。本提案将通过工程和建模先进的细胞系统来研究线粒体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、疾病结局、药物开发和营养的基础知识。代谢生理学的研究也将与教育计划相结合，以激发学生和教育者对生化工程、生物化学和医学的兴趣。为了实现这一目标，我们建议与当地一所高中合作开展一个基于实验室的项目，让学生接触生物工程研究及其在人体生理学中的应用，将酵母代谢与癌症和糖尿病等人类疾病联系起来。此外，还将开发一个基于网络的互动工具，以提高高年级学生和研究人员学习生物化学和代谢的能力。在人类疾病和代谢工程应用的背景下，提出了在真核生物中选择性控制底物利用的新靶点。线粒体与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。尽管在上个世纪对线粒体结构和代谢活动进行了广泛的研究，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）提供了量化低等生物细胞生理的最有效手段，但应用于更复杂的物种因区隔化而变得复杂。研究者开发了一种新的方法来追踪特定亚细胞室的代谢途径，如线粒体和细胞质。在项目的第一个目标中，这项技术将被用来开发一个全面的真核代谢和细胞氧化还原电位的分区模型。细胞工程策略将应用于功能性注释基因/蛋白质运输代谢物进出线粒体。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。这些结果将显著促进对真核细胞代谢的功能理解，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代谢的知识。这一教育计划包括努力(i)强调STEM领域，特别是工程方法对人类健康、工业和环境的影响；（ii）与当地一所高中共同启动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与人体生理学的相关性；（三）开发基于网络的学习工具，作为高级设计课程的一部分，以促进和加快学生和研究人员对代谢途径互联性的学习。这些研究和教育活动也将为本科生、研究生和博士后提供培训机会。该职业奖由CBET部门的生物技术和生化工程项目颁发，由分子和细胞生物学部门的系统和合成生物学项目共同资助。

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.