Polyhydroxylated [60]fullerene binds specifically to functional recognition sites on a monomeric and a dimeric ubiquitin.
Polyhydroxylated [60]fullerene binds specifically to functional recognition sites on a monomeric and a dimeric ubiquitin.
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DOI：
10.1039/c5nr00539f
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发表时间：
2015-04-28
期刊：
影响因子：
6.7
通讯作者：
D'Onofrio M
中科院分区：
文献类型：
--
作者：
Zanzoni S;Ceccon A;Assfalg M;Singh RK;Fushman D;D'Onofrio M
纳米粒子（NPs）在生物医学应用中的使用需要深入了解NPs与生物分子相互作用的机制。与蛋白质结合的纳米颗粒可能会干扰蛋白质-蛋白质相互作用并影响细胞通讯途径，然而纳米颗粒对生物分子识别的影响仍然很难表征。在这方面，特别相关的是NP诱导的蛋白质的功能扰动的研究涉及的关键生化途径的调节。泛素（Ubiquitin，Ub）是一种典型的蛋白质翻译后修饰物，在许多重要的生物学过程中发挥着重要作用。为了有助于了解这种普遍分布的生物大分子和纳米颗粒之间的相互作用，我们研究了多羟基化[60]富勒烯的吸附单体Ub和最小的聚泛素链在体外原子分辨率。站点分辨的化学位移和强度扰动的Ub的NMR信号，连同15 N自旋弛豫速率的变化，交换饱和转移效应，和荧光猝灭的数据是一致的可逆形成的可溶性聚集体纳入富勒醇簇。确定了特异性相互作用表位，与单体和赖氨酸48连接的二聚体Ub中的功能识别位点一致。富勒烯醇似乎针对开放状态的动态结构的二聚体Ub根据构象选择机制。重要的是，蛋白质-NP缔合阻止了多聚泛素链的酶催化合成。我们的研究结果提供了基于实验的蛋白质/富勒烯醇识别的见解，在功能性生物分子通信，包括调节蛋白质周转的影响，并为治疗干预的机会，在UB依赖性细胞通路。
The use of nanoparticles (NPs) in biomedical applications requires an in-depth understanding of the mechanisms by which NPs interact with biomolecules. NPs associating with proteins may interfere with protein-protein interactions and affect cellular communication pathways, however the impact of NPs on biomolecular recognition remains poorly characterized. In this respect, particularly relevant is the study of NP-induced functional perturbations of proteins implicated in the regulation of key biochemical pathways. Ubiquitin (Ub) is a prototypical protein post-translational modifier playing a central role in numerous essential biological processes. To contribute to an understanding of the interactions between this universally distributed biomacromolecule and NPs, we investigated the adsorption of polyhydroxylated [60]fullerene to monomeric Ub and to a minimal polyubiquitin chain in vitro at atomic resolution. Site-resolved chemical shift and intensity perturbations of Ub’s NMR signals, together with 15N spin relaxation rate changes, exchange saturation transfer effects, and fluorescence quenching data were consistent with the reversible formation of soluble aggregates incorporating fullerenol clusters. Specific interaction epitopes were identified, coincident with functional recognition sites in monomeric and lysine48-linked dimeric Ub. Fullerenol appeared to target the open state of the dynamic structure of dimeric Ub according to a conformational selection mechanism. Importantly, protein-NP association prevented enzyme-catalyzed synthesis of polyubiquitin chains. Our findings provide experiment-based insight into protein/fullerenol recognition, with implications in functional biomolecular communication, including regulatory protein turnover, and for the opportunity of therapeutic intervention in Ub-dependent cellular pathways.
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影响因子：
16.6
作者：
Liu, Zhu;Zhang, Wei-Ping;Xing, Qiong;Ren, Xuefeng;Liu, Maili;Tang, Chun
通讯作者：
Tang, Chun
DOI：
10.1039/c39930001784
发表时间：
1993-12-07
影响因子：
--
作者：
LI, J;TAKEUCHI, A;KITAZAWA, K
通讯作者：
KITAZAWA, K
影响因子：
17.1
作者：
Calvaresi, Matteo;Arnesano, Fabio;Zerbetto, Francesco
通讯作者：
Zerbetto, Francesco
影响因子：
5.6
作者：
Haririnia, Aydin;D'Onofrio, Mariapina;Fushman, David
通讯作者：
Fushman, David
DOI：
10.1073/pnas.0701250104
发表时间：
2007-05-22
影响因子：
11.1
作者：
Linse, Sara;Cabaleiro-Lago, Celia;Dawson, Kenneth A.
通讯作者：
Dawson, Kenneth A.