CAREER: Quantification of Prion Protein Sorption to Soil Components

职业：朊病毒蛋白对土壤成分吸附的定量

• 批准号: 0547484
• 负责人: Joel Pedersen
• 金额: $ 40万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2013-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0547484&HistoricalAwards=false
• 关键词: CAREER; Quantification; Prion; Protein; Sorption

Pedersen 0547484The main objectives of this NSF CAREER proposal are (1) to develop a highly sensitive, quantitative method for prion protein measurement, and (2) to quantitatively describe the association of prion proteins with surface-reactive soil minerals and to examine the factors influencing attachment to and detachment from these particles. Quantitative analytical methods for prion proteins are currently lacking. The PI therefore intends to exploit recent advances in proteomics to develop a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for prion protein quantitation based on a signature peptide approach. The second objective will be addressed in a series of batch attachment and detachment experiments using the newly developed LC-MS/MS method for quantitation. The association of prion proteins to clay minerals and metal oxides will be examined as a function of pH to determine the influence of prion and mineral surface charge on attachment. Reversibility of prion protein attachment due to shifts in pH and the presence of surfactants and dissolved organic matter will be investigated in batch detachment experiments. The importance of electrostatic and hydrophobic interactions between prions and mineral surfaces will be explicitly examined. Experimental results will be interpreted within the framework of extended Derjaguin-Landau-Verwey-Overbeek theory. The PI will use atomic force microscopy to measure interaction forces between prion protein and a subset of mineral surfaces. From these experiments, the PI will derive sticking efficiencies to facilitate prediction of prion transport in porous media.Broader impacts: The LC-MS method is expected to have utility well beyond the environmental fate studies outlined in this proposal. The method has the potential to significantly impact medical and veterinary research and the diagnosis of transmissible spongiform encephalopathies. Improved understanding of prion attachment to soil components will naturally lead to the development of engineering applications based on our findings. The educational component of this career development plan focuses on integrating research and instruction and fostering increased interdisciplinarity in environmental engineering and science. The PI will develop a new graduate course on environmental colloid chemistry and a short course for practicing environmental professionals on the treatment and disposal of biological threat agents. Broader impacts will also be realized through enhanced K-12 education. The PI will extend an existing collaboration with a middle school science teacher to design, implement and evaluate an instructional unit on environmental engineering and science organized around inquiry- and discovery-based learning exercises. In addition to publishing research findings in leading journals, information about prions in the environment will be disseminated to a wider audience through a "display" in the UW Madison Virtual Museum of Minerals and Molecules. During the duration of the project, the PI will provide opportunities for undergraduate students to participate in research, with special consideration given to underrepresented minority applicants. The PI will leverage existing resources to reach his educational objectives such as UW NSF-funded Delta Program in Research, Teaching and Learning to provide opportunities for graduate students to develop skills in classroom teaching, preparation of instructional materials, informal education, teaching to diverse student audiences, teaching with technology and internships.

PERDERSEN 0547484这份NSF职业计划的主要目标是(1)开发一种高灵敏度的定量测量普恩蛋白的方法，以及(2)定量描述普恩蛋白与土壤表面活性矿物的关联，并研究影响这些颗粒附着和脱离的因素。目前还缺乏对病毒蛋白进行定量分析的方法。因此，PI打算利用蛋白质组学的最新进展，基于标志性多肽方法，开发一种用于Pron蛋白定量的LC-MS/MS方法。第二个目标将在一系列批量附着和分离实验中使用新开发的LC-MS/MS定量方法来解决。Pron蛋白质与粘土矿物和金属氧化物的结合将作为pH的函数来检测，以确定Prion和矿物表面电荷对附着的影响。在分批分离实验中，将研究由于pH变化以及表面活性剂和溶解有机物的存在而引起的蛋白附着的可逆性。我们将明确研究Pron和矿物表面之间的静电和疏水相互作用的重要性。实验结果将在扩展的Derjaguin-Landau-Verwey-Overbeek理论框架内解释。PI将使用原子力显微镜来测量Pron蛋白质和矿物表面的子集之间的相互作用力。从这些实验中，等电点将获得粘贴效率，以便于预测普恩在多孔介质中的传输。广泛的影响：LC-MS方法预计将具有远远超过本提案中概述的环境命运研究的实用价值。该方法有可能对医学和兽医研究以及传染性海绵状脑病的诊断产生重大影响。根据我们的发现，对Pron附着在土壤成分上的了解的提高自然会导致工程应用的发展。这一职业发展计划的教育部分侧重于将研究和教学结合起来，并促进环境工程和科学方面更多的跨学科。该协会将开设一门新的环境胶体化学研究生课程，并为环境专业人员开设一门有关生物威胁物质处理和处置的短期课程。还将通过加强K-12教育实现更广泛的影响。PI将扩展与一名中学科学教师的现有合作，以设计、实施和评估围绕探究性和发现性学习练习组织的环境工程和科学教学单元。除了在主要期刊上发表研究成果外，有关环境中的普恩病毒的信息还将通过威斯康星州麦迪逊大学矿物和分子虚拟博物馆的“展示”向更广泛的受众传播。在项目期间，国际学生协会将为本科生提供参与研究的机会，并特别考虑代表人数不足的少数族裔申请者。PI将利用现有资源实现他的教育目标，如UW NSF资助的Delta研究、教学和学习计划，为研究生提供在课堂教学、教材准备、非正式教育、面向不同学生受众的教学、技术教学和实习方面的技能发展机会。