Ligand-Binding Cooperativity Effects in Polymer-Protein Conjugation.
Ligand-Binding Cooperativity Effects in Polymer-Protein Conjugation.
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聚合物-蛋白质缀合中的配体结合协同效应
DOI：
10.1021/acs.biomac.9b00016
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发表时间：
2019
影响因子：
6.2
通讯作者：
Dariush Hinderberger
中科院分区：
文献类型：
--
作者：
Jörg Reichenwallner;Anja Thomas;Tobias Steinbach;Jana Eisermann;Christian E. H. Schmelzer;Frederik Wurm;Dariush Hinderberger
我们提出了一个电子顺磁共振（EPR）光谱表征的结构和动力学效应，源于翻译后修饰的牛血清白蛋白（BSA），一个既定的模型系统聚合物-蛋白质结合。除了这种系统的典型药物输送和生物相容性方面，我们说明了改变内部动力学的原因，因此在BSA蛋白质核心的配体结合方面的功能。吸收顺磁性脂肪酸衍生物16-doxyl硬脂酸由几个BSA为基础的方酸大分子引发剂和聚合物-蛋白质缀合物的EPR光谱研究，辅助动态光散射（DLS）和zeta电位测量。共轭物接枝从寡（乙二醇）甲基醚甲基丙烯酸酯（OEGMA），形成一个整体的核-壳状结构。结果发现，配体结合和相关的参数，如结合亲和力，协同性，和BSA的结合位点的数量的变化与表面改性的程度急剧。在处理BSA的过程中，配体也改变了它们对单个结合位点的偏好，如从双电子电子共振（DEER）实验中它们的空间排列的比较视图所观察到的。蛋白质附着的聚合物构成扩散屏障，显著阻碍配体摄取。此外，zeta电位（Zeta电位）与蛋白质大分子引发剂的表面改性程度呈线性下降，并且可以估计缀合物中聚合物层的有效介电常数。所有这些都表明，BSA中的配体摄取特性可以通过这种翻译后修饰（PTM）的程度和性质进行微调。我们表明，EPR光谱是合适的量化这些微妙的PTM为基础的功能效应从自组装基板和配体。
We present an electron paramagnetic resonance (EPR) spectroscopic characterization of structural and dynamic effects that stem from post-translational modifications of bovine serum albumin (BSA), an established model system for polymer–protein conjugation. Beyond the typical drug delivery and biocompatibility aspect of such systems, we illustrate the causes that alter internal dynamics and therefore functionality in terms of ligand-binding to the BSA protein core. Uptake of the paramagnetic fatty acid derivative 16-doxyl stearic acid by several BSA-based squaric acid macroinitiators and polymer–protein conjugates was studied by EPR spectroscopy, aided by dynamic light scattering (DLS) and zeta potential measurements. The conjugates were grafted from oligo(ethylene glycol) methyl ether methacrylate (OEGMA), forming an overall core–shell-like structure. It is found that ligand-binding and associated parameters such as binding affinity, cooperativity, and the number of binding sites of BSA change drastically with the extent of surface modification. In the course of processing BSA, the ligands also change their preference for individual binding sites, as observed from a comparative view of their spatial alignments in double electron electron resonance (DEER) experiments. The protein-attached polymers constitute a diffusion barrier that significantly hamper ligand uptake. Moreover, zeta potentials (ζ) decrease linearly with the degree of surface modification in protein macroinitiators and an effective dielectric constant can be estimated for the polymer layer in the conjugates. All this suggests that ligand uptake characteristics in BSA can be fine-tuned by the extent and nature of such post-translational modifications (PTMs). We show that EPR spectroscopy is suitable for quantifying these subtle PTM-based functional effects from self-assembly of substrate and ligand.
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硬脂酸自旋标签与人血清白蛋白的结合：一项 EPR 研究。
DOI：
10.1021/jp5068928
发表时间：
2014
期刊：
The journal of physical chemistry. B
影响因子：
--
作者：
A. Pavićević;A. Popović;M. Mojović;Snežana V Šušnjar;G. Bačić
通讯作者：
G. Bačić
影响因子：
15
作者：
KARUSH, F
通讯作者：
KARUSH, F
影响因子：
7.4
作者：
Bonanata, Jenner;Turell, Lucia;Laura Coitino, E.
通讯作者：
Laura Coitino, E.
影响因子：
3.5
作者：
DOLHOFER, R;WIELAND, OH
通讯作者：
WIELAND, OH
影响因子：
6.2
作者：
Morgenstern, Josefine;Alvaradejo, Gabriela Gil;Hubbuch, Juergen
通讯作者：
Hubbuch, Juergen