Carbon Sequestration: Tracing the Source and Fate of Marine Biopolymers using Proteomics and Antibody Display Technology

溶解生物高分子材料的化学特性和动力学是海洋学中一个长期存在的问题。然而，由于先前缺乏研究溶解生物聚合物的技术，我们才刚刚开始了解分子水平上的来源、组成和反应性。在这项研究中，系统生物学研究所的科学家将利用高通量蛋白质组学和抗体显示技术开发用于生物聚合物研究的创新工具（生物传感器）。而不是仅仅检查水解后的单个氨基酸，这抹去了蛋白质的化学历史和生物学与海洋化学之间的联系，蛋白质组学的应用来表征两种模式浮游植物的分泌蛋白质组学将促进我们在分子水平上对这些化合物的组成和反应性的理解。由于分泌蛋白质组被认为是保守的，了解这两个重要生态物种的蛋白质组应该有助于其他浮游植物分泌蛋白质组的表征。高亲和力生物传感器（来自噬菌体的单克隆抗体）可以识别并结合特定的分子片段，这将允许在复杂的异质样品中对低浓度的化学物质进行鉴定和定量。最终，这项技术将允许浮游植物蛋白质的原位跟踪，它们的生物合成，转化和在不浓缩的海水中的降解。就更广泛的影响而言，结合浮游植物和蓝藻释放蛋白质的直接监测，以及利用单克隆探针跟踪浮游植物和蓝藻蛋白质，应该可以通过了解浮游植物和蓝藻释放的蛋白质进入整个溶解有机物库的动态和周转，将生物生产和消费与海洋化学联系起来。此外，这将使我们更好地了解它们在全球碳循环中的作用，并将大大提高我们对海洋地球化学和海洋生态学的认识。作为这项研究的一部分，本科生将接触到这些新技术。pi将在地方和区域学区的科学教育活动中发挥积极作用。

ABSTRACTOCE-0452563The chemical identity and dynamics of dissolved biopolymeric material is a standing question in oceanography. However, due to a prior dearth of techniques to study dissolved biopolymers, we are only beginning to understand the source, composition and reactivity at the molecular level. In this research, scientists from the Institute of Systems Biology will develop innovative tools (biosensors) for biopolymer research using high throughput proteomics and antibody display technology. Rather than solely examining single amino acids after hydrolysis, which erases the chemical-history of the proteins and the link between biology and ocean chemistry, the application of proteomics to characterize the secretory proteome of two model phytoplankton species will advance our understanding of the composition and reactivity of these compounds at the molecular level. Because secretory proteomes are thought to be conservative, knowledge of the proteome for these two ecologically important species should aid in the characterization of other phytoplankton secretory proteomes. High affinity biosensors (monoclonal antibodies from phage) that recognize and bind to specific molecular moieties will allow the identification and quantification of chemical species at low concentrations in complex heterogeneous samples. Ultimately this technology will allow in situ tracking of phytoplankton proteins, their biosynthesis, transformation, and degradation in unconcentrated seawater. In terms of broader impacts, combining the direct monitoring of proteins released by phytoplankton and cyanobacteria and tracking of phytoplankton and cyanobacterial proteins with monoclonal probes should allow the linking of biological production and consumption to ocean chemistry through understanding the dynamics and turnover of proteins released by phytoplankton and cyanobacteria into the overall dissolved organic matter pool. Furthermore, this will lead to improved knowledge of their role in global carbon cycling and will greatly advance our knowledge of marine geochemistry and marine ecology. Undergraduate students will be exposed to these new techniques as part of this study. The PIs will play active roles in local and regional school district science educational initiatives.