RUI: Chemically Modified Enzymes to Control Adsorption on Gold Nanoparticles for Enhanced Structure/Function

RUI：化学修饰酶控制金纳米粒子上的吸附以增强结构/功能

基本信息

批准号：
2203740
负责人：
Jeremy Driskell
金额：
$ 37.69万
依托单位：
Board of Trustees of Illinois State University
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=2203740&HistoricalAwards=false
关键词：
RUI Chemically Modified Enzymes Control

项目摘要

With support from the Macromolecular, Supramolecular and Nanochemistry Program in the Division of Chemistry, Professor Jeremy Driskell of Illinois State University is combining chemical synthesis and advanced chemical analysis tools to study how proteins adsorb onto gold nanoparticles and maintain or enhance their biological function. The study identifies key interactions between the protein and nanoparticle that can be exploited to prevent protein unfolding and to facilitate controlled release from the nanoparticle surface. Professor Driskell and his students are chemically modifying proteins to install chemical anchors with high affinity for the gold nanoparticles and measuring the stability and biological function of the adsorbed protein layer. Their discoveries could lead to predictive design parameters to form robust and highly functional protein-nanoparticle probes and broadly impact modern biosensing, medical imaging, drug delivery, and biocatalysis. This project provides support for undergraduate students to participate in an immersive research experience to learn advanced analytical techniques and gain an appreciation for a multidisciplinary approach to problem solving. Additionally, in collaboration with the Illinois Research Academy, Dr. Driskell and his supported undergraduate students provide high school students with an intense, yet supervised, research experience. A detailed understanding of the protein-nanoparticle interface is critical to mitigate structural changes that negatively impact the function of the adsorbed protein and to leverage stabilizing interactions that enhance protein function. Surface accessible thiols on a protein are hypothesized to be primarily responsible for the adsorption onto gold nanoparticles and the formation of a hard corona; thus, the ability to precisely control the number of protein thiols can be exploited to optimize bioconjugate function. A synthetic strategy is proposed to install thiol functional groups on a series of enzymes. The reaction conditions are optimized to precisely control the number of thiols, and monitored by high-resolution mass spectrometry, zeta potential, and Ellman’s reagent. Adsorption affinity, quantitatively assessed using nanoparticle tracking analysis, and protein exchange rate, measured via a competitive protein binding, is correlated with the number of surface accessible thiols presented by the protein. Additionally, the structure and function of the free and nanoparticle-immobilized protein is compared to identify any relationship between protein thiolation and protein structure/function upon adsorption to a nanoparticle. Successful completion of this project drives the novel design of highly active and stable protein-AuNP conjugates that is critically needed to advance bioconjugate-enabled platform technologies.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系高分子、超分子和纳米化学项目的支持下，伊利诺伊州立大学的 Jeremy Driskell 教授正在将化学合成和先进的化学分析工具结合起来，研究蛋白质如何吸附到金纳米颗粒上并维持或增强其生物功能。 Driskell 教授和他的学生正在利用蛋白质和纳米粒子之间的相互作用来阻止蛋白质展开并促进纳米粒子表面的受控释放。他们的发现可以产生对金纳米颗粒具有高亲和力的锚并测量吸附蛋白质层的稳定性和生物功能，从而可以形成预测设计参数，以形成坚固且功能强大的蛋白质纳米颗粒探针，并广泛影响现代生物传感、医学成像、药物输送。该项目为本科生提供参与沉浸式研究体验的支持，以学习先进的分析技术并获得对解决问题的多学科方法的欣赏。 Driskell 和他支持的本科生为高中生提供了深入但受监督的研究经验，这对于减轻对吸附蛋白质功能产生负面影响的结构变化以及利用稳定的相互作用至关重要。蛋白质表面可接近的硫醇被捕获，主要负责金纳米颗粒的吸附和硬电晕的形成，因此，可以利用精确控制蛋白质硫醇数量的能力；提出了一种在一系列酶上安装硫醇官能团的合成策略，以精确控制硫醇的数量，并通过高分辨率质谱、zeta 电位和埃尔曼试剂进行监测。使用纳米颗粒跟踪分析定量评估的亲和力和通过竞争性蛋白质结合测量的蛋白质交换率与蛋白质呈现的表面可接触硫醇的数量相关。比较游离和纳米颗粒固定的蛋白质，以确定蛋白质硫醇化和吸附到纳米颗粒后的蛋白质结构/功能之间的关系。该项目的成功完成推动了高活性和稳定的蛋白质-AuNP 缀合物的新颖设计，这是推进所急需的。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查标准进行评估，被认为值得支持。