Probing Dynamics and Structure of the Nanoparticle Protein Corona to Understand Its Impacts on Environmental Health and Safety

探测纳米颗粒蛋白电晕的动力学和结构，以了解其对环境健康和安全的影响

• 批准号: 1438634
• 负责人: Stephan Link
• 金额: $ 30万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1438634&HistoricalAwards=false
• 关键词: Probing; Dynamics; Structure; Nanoparticle; Protein

PI: Link, Stephan Proposal Number: 1438634 Institution: William Marsh Rice UniversityTitle: Probing Dynamics and Structure of the Nanoparticle Protein Corona to Understand Its Impacts on Environmental Health and Safety The properties of nanoparticles (NPs) in biological fluids differ from their behavior in water due to the association of different proteins to the NP surface, forming a "protein corona" around the NPs. These proteins bind strongly to the NPs and affect the NPs' interactions with their environment. The goal of this proposal is to quantify the dynamics of proteins binding to NPs and to determine how the proteins may change their structure upon binding. The project will investigate model proteins, as well as proteins in blood plasma. The inclusion of experiments with whole plasma is an attractive feature of this proposed work, as whole plasma reflects the complexity of real-world conditions. The results of this research will be important for the understanding of the fate of nanoparticles, particularly, the bio-accumulation of nanoparticles in the food chain. In addition, understanding the dynamics of protein coating of NPs can be useful for the development of approaches that intentionally coat NPs with proteins in order to prevent other proteins from binding as a strategy to ensure that the NPs do not cause adverse effects on the environment. The PI and Co-PI are both active in outreach to the K-12 community and mentoring of undergraduate and graduate students. While it is believed that the corona changes over time, as initially the most abundant proteins bind, but are then replaced with proteins that have the largest affinity, very little is known about the time scales of this dynamic exchange among different proteins. The goal of this proposal is to quantify the time-scales of protein-binding to NPs and to determine binding-induced structural changes of the proteins. Correlation spectroscopy will be used to determine the equilibrium protein-NP binding constant and number of bound proteins. The approach will be extended to measure competitive binding among different proteins to the same or possibly different nanoparticles. For protein folding/unfolding on NPs, the PIs will use CD spectroscopy on individual nanoparticles (with the goal of achieving single protein detection levels) and smFRET (single molecule fluorescence resonance energy transfer) microscopy of donor-acceptor labeled proteins. The investigators have unique and complementary strong expertise in the proposed area of research.

主要研究者：Link，Stephan提案编号：1438634机构：威廉·马什赖斯大学标题：探测动力学和纳米颗粒蛋白质电晕的结构，以了解其对环境健康和安全的影响纳米颗粒（NP）在生物流体中的性质不同于它们在水中的行为，这是由于不同的蛋白质与NP表面的缔合，在NP周围形成“蛋白质电晕”。这些蛋白质与纳米颗粒强烈结合，并影响纳米颗粒与其环境的相互作用。该提案的目标是量化蛋白质与NP结合的动力学，并确定蛋白质在结合后如何改变其结构。该项目将研究模型蛋白质以及血浆中的蛋白质。包含整个等离子体的实验是这项拟议工作的一个有吸引力的特点，因为整个等离子体反映了现实世界条件的复杂性。这项研究的结果对于理解纳米颗粒的命运，特别是纳米颗粒在食物链中的生物积累将是重要的。此外，了解NP的蛋白质包被的动力学可以用于开发有意用蛋白质包被NP以防止其他蛋白质结合的方法，作为确保NP不会对环境造成不利影响的策略。PI和Co-PI都积极参与K-12社区的外联活动，并指导本科生和研究生。虽然人们认为冠随着时间的推移而变化，因为最初最丰富的蛋白质结合，但随后被具有最大亲和力的蛋白质取代，但对不同蛋白质之间这种动态交换的时间尺度知之甚少。该提案的目标是量化蛋白质结合到NP的时间尺度，并确定结合诱导的蛋白质结构变化。相关光谱将用于确定平衡蛋白质-NP结合常数和结合蛋白质的数量。该方法将扩展到测量不同蛋白质与相同或可能不同的纳米颗粒之间的竞争性结合。对于纳米颗粒上的蛋白质折叠/解折叠，PI将使用单个纳米颗粒上的CD光谱（目标是实现单个蛋白质检测水平）和供体-受体标记蛋白质的smFRET（单分子荧光共振能量转移）显微镜。研究人员在拟议的研究领域拥有独特和互补的强大专业知识。