EAGER: Marine biopolymers as tracers of major biogeochemical processes: Using proteomics and antibody-sensor technology

EAGER：海洋生物聚合物作为主要生物地球化学过程的示踪剂：使用蛋白质组学和抗体传感器技术

• 批准号: 1219537
• 负责人: Monica Orellana
• 金额: $ 29.98万
• 依托单位: University of Washington
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-15 至 2015-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1219537&HistoricalAwards=false
• 关键词: EAGER; Marine; biopolymers; tracers; major

Recent advances in proteomics, biomarkers and biosensor technology sciences enable new approaches to understanding major biogeochemical processes. This project will examine the physicochemical reactivity of a model protein "RuBisCO" in seawater, and will quantify RuBisCO along ocean transect Line P (48°39.0' N, 126°40.0' W to 50°00' N, 145°00' W) in the North Pacific Ocean. The project will use two independent methods that complement and validate each other: immune-sensors and multiple-reaction monitoring (MRM) mass spectrometry.Intellectual Merit: Chemical analyses have shown that a significant fraction of dissolved organic matter (DOM) in the ocean is in the form of proteins. Proteins are a rich source of biological information and their amino acid sequence provides a direct link to the coding DNA of an organism. Identification of proteins in DOM opens a window to understanding the complex sources and dynamics of biopolymeric material. Furthermore, proteins represent the catalytic potential and reactivity of an organism and, collectively, of an ecosystem. Most importantly, the information also provides a direct link between biological and phylogenetic presence and biogeochemistry. The distributions of proteins that are specific to major processes (e.g., RuBisCO to carbon fixation, nitrogenase to diazotrophy) can be used to infer information that has been hidden until now. The PIs have recently demonstrated the deep distribution of RubisCO in the North Pacific and discovered this enzyme to be at high concentrations throughout the water column (to depths 3000 m) underlying highly productive equatorial and subpolar systems, and low concentrations under the oligotrophic subtropical gyre. This single protein represents ~2% of the largely unidentified dissolved organic nitrogen pool at depths 1000 m. The deep distribution of RuBisCO shows that hydrographic fronts in the surface ocean affect the distribution of recently-produced organic matter thousands of meters below the ocean surface, and the enzyme traces the transport of deep organic matter by the deep ocean circulation away from regions of input. These findings suggest that the identification and quantification of additional biomarker proteins will provide a powerful approach to understanding the associated biogeochemical pathways. While potentially transformative, the protocols need to be explored and validated. In this study , the PIs will compare and validate their previous findings by understanding the physicochemical reactivity of RuBisCO (and therefore other proteins) in seawater samples in order to fully interpret and exploit the information held in protein distributions; and they will quantify RuBisCO at Line P in fresh samples using two independent and different methods that complement and validate each other: immunosensors and multiple-reaction monitoring (MRM) mass spectrometry in a coastal to open ocean transect.Broader Impacts:Proteomics is in its infancy in the aquatic sciences and this research will help drive its development.Merging the sciences of proteomics with biogeochemistry will leverage both. The Sorcerer II Global Ocean Sampling (GOS) expedition predicted more than six million proteins in the GOS database, almost twice the number of proteins present in the current databases, adding an enormous diversity to the known protein families. Using immunosensors coupled with S/MRM proteomics will provide information on the cycling of RuBisCO as an actual biopolymers in the DOM pool. Current methods use hydrolysis which solely examines single amino acids but erases the chemical history of the proteins and the link between biology, phylogenetic presence, and biogeochemistry. Protein immunosensors will allow in situ tracking of marine proteins, their biosynthesis, transformation, and degradation in unconcentrated seawater. Furthermore, understanding the formation, degradation, and preservation of proteins in the DOC/DON (dissolved organic carbon/nitrogen) pools will elucidate their role in global carbon and nitrogen cycling and will greatly advance our knowledge of marine biogeochemistry.

蛋白质组学、生物标志物和生物传感器技术科学的最新进展为理解主要的生物地球化学过程提供了新的途径。该项目将研究模型蛋白质“Rubisco”在海水中的物理化学反应活性，并将沿着北太平洋的海洋横断面P线(48°39.0‘N，126°40.0’W到50°00‘N，145°00’W)对Rubisco进行量化。该项目将使用两种相互补充和验证的独立方法：免疫传感器和多反应监测(MRM)质谱仪。智力优势：化学分析表明，海洋中溶解的有机物(DOM)中有很大一部分是以蛋白质的形式存在的。蛋白质是丰富的生物信息来源，它们的氨基酸序列提供了与生物体编码DNA的直接联系。DOM中蛋白质的鉴定为理解生物聚合物材料的复杂来源和动力学打开了一扇窗。此外，蛋白质代表了有机体和生态系统的催化潜力和反应能力。最重要的是，这些信息还提供了生物和系统发育存在与生物地球化学之间的直接联系。主要过程特有的蛋白质的分布(例如，Rubisco到固碳，固氮酶到重氮化)可以用来推断到目前为止一直被隐藏的信息。PIS最近证实了Rubisco在北太平洋的深层分布，并发现这种酶在高生产力的赤道和亚极地系统下方的水柱(深达3000米)中浓度很高，而在少营养的亚热带环流下浓度很低。这一单一蛋白质约占1000米深处大部分未知的溶解有机氮库的2%。Rubisco的深层分布表明，表层海洋中的水文锋面影响着海面下数千米处新近产生的有机质的分布，该酶跟踪深海环流向输入区域以外的深层有机质的运输。这些发现表明，鉴定和量化额外的生物标志物蛋白将为理解相关的生物地球化学途径提供一个强有力的途径。虽然这些协议具有潜在的变革性，但仍需进行探索和验证。在这项研究中，PI将通过了解海水样品中Rubisco(以及其他蛋白质)的物理化学反应活性来比较和验证他们之前的发现，以便充分解释和利用蛋白质分布中包含的信息；他们将使用两种相互补充和验证的独立和不同的方法来量化P线新鲜样品中的Rubisco：免疫传感器和多反应监测(MRM)质谱仪，在沿海到开阔的海洋横断面上。广泛的影响：蛋白质组学在水产科学中处于初级阶段，这项研究将有助于推动其发展。蛋白质组学科学与生物地球化学的融合将利用两者。Magcerer II全球海洋采样(GOS)探险队预测GOS数据库中有600多万种蛋白质，几乎是目前数据库中蛋白质数量的两倍，为已知的蛋白质家族增加了巨大的多样性。将免疫传感器与S/MRM蛋白质组学结合使用，将提供有关Rubisco作为DOM池中的实际生物聚合物的循环信息。目前的方法使用仅检查单一氨基酸的水解法，但删除了蛋白质的化学史以及生物学、系统发育存在和生物地球化学之间的联系。蛋白质免疫传感器将允许原位跟踪海洋蛋白质及其在非浓缩海水中的生物合成、转化和降解。此外，了解DOC/DON(溶解有机碳/氮)池中蛋白质的形成、降解和保存将有助于阐明它们在全球碳氮循环中的作用，并将极大地促进我们对海洋生物地球化学的了解。