Perera Y Randika, South Taylor M, Hughes Alex C, Parkhurst Ashlyn N, Williams Olivia C, Davidson Mackenzie B, Wilks Chloe A, Mlsna Debra A, Fitzkee Nicholas C
Mississippi State University, Mississippi State, Mississippi.
J Chem Educ. 2020 Mar 10;97(3):820-824. doi: 10.1021/acs.jchemed.9b00625. Epub 2020 Jan 21.
A simple one-dimensional H NMR experiment that quantifies protein bound to gold nanoparticles has been developed for upper-division biochemistry and physical chemistry students. This laboratory experiment teaches the basics of NMR techniques, which is a highly effective tool in protein studies and supports students to understand the concepts of NMR spectroscopy and nanoparticle-protein interactions. Understanding the interactions of gold nanoparticles (AuNPs) with biological macromolecules is becoming increasingly important as interest in the clinical use of nanoparticles has been on the rise. Applications in drug delivery, biosensing, diagnostics, and enhanced imaging are all tangible possibilities with a better understanding of AuNP-protein interactions. The ability to use AuNPs as biosensors for drug delivery methods in cellular uptake is dependent on the amount of protein that is able to bind to the surface of the nanoparticle. This laboratory experiment solidifies concepts such as quantitative NMR spectroscopy while reinforcing precision laboratory titrations. Students learn how H proton NMR spectra can be used to measure free protein in solution and protein bound to AuNPs. A simple formula is used to determine the binding capacity of the nanoparticle. This analysis helps students to understand the impact of nanoparticle-protein interactions, and it allows them to conceptualize macromolecular binding using NMR spectroscopy.
我们为高年级生物化学和物理化学专业的学生开发了一种简单的一维氢核磁共振（¹H NMR）实验，用于量化与金纳米颗粒结合的蛋白质。这个实验室实验教授核磁共振技术的基础知识，该技术在蛋白质研究中是一种非常有效的工具，有助于学生理解核磁共振光谱学和纳米颗粒 - 蛋白质相互作用的概念。随着对纳米颗粒临床应用的兴趣日益增加，了解金纳米颗粒（AuNP）与生物大分子的相互作用变得越来越重要。更好地理解AuNP - 蛋白质相互作用后，在药物递送、生物传感、诊断和增强成像等方面的应用都切实可行。将AuNP用作细胞摄取中药物递送方法的生物传感器的能力取决于能够结合到纳米颗粒表面的蛋白质数量。这个实验室实验巩固了定量核磁共振光谱学等概念，同时强化了精确的实验室滴定操作。学生们学习如何利用¹H质子核磁共振光谱来测量溶液中的游离蛋白质以及与AuNP结合的蛋白质。使用一个简单的公式来确定纳米颗粒的结合能力。这种分析有助于学生理解纳米颗粒 - 蛋白质相互作用的影响，并使他们能够利用核磁共振光谱学来概念化大分子结合。
