Fluorescamine Labeling for Assessment of Protein Conformational Change and Binding Affinity in Protein-Nanoparticle Interaction.
Fluorescamine Labeling for Assessment of Protein Conformational Change and Binding Affinity in Protein-Nanoparticle Interaction.
复制标题
DOI：
10.1021/acs.analchem.7b02810
复制
发表时间：
2017-11-21
影响因子：
7.4
通讯作者：
Zhong W
中科院分区：
文献类型：
--
作者：
Duan Y;Liu Y;Shen W;Zhong W
蛋白质吸附改变了纳米颗粒（NPs）的“生物学特性”，并可能影响生物系统对入侵的NPs的反应。研究蛋白质- np相互作用可以帮助理解np的物理化学性质如何影响相互作用，从而潜在地指导生物医学或其他应用中更安全、更有效的np的设计。蛋白质与NPs之间的结合亲和力以及蛋白质与NPs结合后构象变化的发生是需要学习的两个重要方面，但目前很少有方法可以同时以简单的方式评估这两个方面。本研究证明，本团队开发的荧光胺标记方法不仅能够标记暴露在蛋白质表面的伯胺，从而揭示蛋白质被NPs吸附后的构象变化，而且还可以用于测量蛋白质的结合亲和力。通过筛选大量蛋白质与四种NPs之间的相互作用，本研究还发现蛋白质在NPs上的吸附可能受到结构柔韧性的强烈影响。具有高结构柔韧性的蛋白质在与聚苯乙烯NPs结合时发生了高度的构象变化，这可能会影响蛋白质的功能。总的来说，我们证明了我们的分析是一种快速、简单和高通量的工具，可以揭示对蛋白质活性的潜在影响，并评估蛋白质- np结合的强度。
Protein adsorption alters the “biological identity” of the nanoparticles (NPs) and could affect how biosystems respond to the invading NPs. Study of protein-NP interaction can help understand how the physicochemical properties of NPs impact the interaction and thus potentially guide the design of safer and more effective NPs for biomedical or other applications. Binding affinity between proteins and NPs, and the occurrence of protein conformational change upon binding to NPs are two important aspects to be learned, but few methods are currently available to assess both simultaneously in a simple way. Herein, we demonstrated that the fluorescamine labeling method developed in our group could not only reveal protein conformational change upon adsorption to NPs, owing to its capability to label the primary amines exposed on protein surface, but also it could be applied to measure the binding affinity. By screening the interaction between a large number of proteins and four types of NPs, the present study also revealed that protein adsorption onto NPs could be strongly affected by structure flexibility. The proteins with high structure flexibility experienced high degrees of conformation change when binding to the polystyrene NPs, which could potentially influence protein function. Overall, we demonstrate that our assay is a quick, simple, and high-throughput tool to reveal potential impacts on protein activity and evaluate the strength of protein-NP binding.
登录
查看更多内容
影响因子：
3.9
作者：
Gebauer, Julia S.;Malissek, Marcelina;Treuel, Lennart
通讯作者：
Treuel, Lennart
影响因子：
2.9
作者：
BJELLQVIST, B;HUGHES, GJ;HOCHSTRASSER, D
通讯作者：
HOCHSTRASSER, D
影响因子：
3.9
作者：
DE BERNARDO, S;WEIGELE, M;UDENFRIEND, S
通讯作者：
UDENFRIEND, S
影响因子：
17.1
作者：
Albanese, Alexandre;Walkey, Carl D.;Chan, Warren C. W.
通讯作者：
Chan, Warren C. W.
影响因子：
17.1
作者：
Casals, Eudald;Pfaller, Tobias;Puntes, Victor
通讯作者：
Puntes, Victor