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荧光蛋白纳米颗粒:细胞氧化损伤的合成与识别。

Fluorescent protein nanoparticles: Synthesis and recognition of cellular oxidation damage.

机构信息

State Key Laboratory of Virology & Key Laboratory of Analytical Chemistry for Biology and Medicine (MOE), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, PR China.

State Key Laboratory of Virology & Key Laboratory of Analytical Chemistry for Biology and Medicine (MOE), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, PR China; Guangxi Key Laboratory of Natural Polymer Chemistry, College of Chemistry and Material Sciences, Nanning Normal University, Nanning 530001, PR China; Key Laboratory of Coal Conversion and Carbon Materials of Hubei Province, College of Chemistry and Chemical Engineering, Wuhan University of Science and Technology, Wuhan 430081, PR China.

出版信息

Colloids Surf B Biointerfaces. 2019 May 1;177:219-227. doi: 10.1016/j.colsurfb.2019.01.065. Epub 2019 Feb 1.

Abstract

Intracellular reactive oxygen species (ROS) generation are associated with many diseases. Lots of studies focus on the detection of intracellular ROS by small fluorescent molecules. However, ROS recognized by biocompatible nanoparticles are relatively less reported. It is widely known that albumin-based nanomaterials possess unique advantages in biomedical applications because they are biodegradable and biocompatible. Herein, fluorescent protein nanoparticles (PNPs) were prepared using BSA as a starting material without introducing extra fluorescent molecules. The blue fluorescent PNPs were well characterized by FL, FTIR, CD, TEM, DLS, etc. It was revealed that the PNPs exhibited two types of emissive centers through FL spectra and the fluorescence lifetimes. Further mechanism study indicated that the fluorescence of the PNPs was mainly derived from three kinds of aromatic amino acids, namely tryptophan, tyrosine and phenylalanine. Moreover, the fluorescence properties of the PNPs were tightly related to pH. The PNPs displayed excellent stabilities under harsh conditions as well as physiological conditions. In addition, the PNPs (200 μg/mL) were nontoxic to HeLa and GES-1 cell lines, showing good biocompatibility. The cellular uptake of PNPs was occurred only when the cells were stressed with glucose oxidase or HO, thereafter the bright blue fluorescence was observed, indicating that it could be utilized for the recognition of cellular oxidation damage. These findings will offer novel clues for the future synthesis of even brighter protein nanoparticles and their biomedical applications.

摘要

细胞内活性氧(ROS)的产生与许多疾病有关。许多研究都集中在通过小分子荧光探针来检测细胞内 ROS。然而,相对较少有研究报道生物相容性纳米颗粒所识别的 ROS。众所周知,基于白蛋白的纳米材料在生物医学应用中具有独特的优势,因为它们可生物降解且生物相容性良好。在此,我们使用 BSA 作为起始原料,而无需引入额外的荧光分子,制备了荧光蛋白纳米颗粒(PNPs)。通过 FL、FTIR、CD、TEM、DLS 等对蓝色荧光 PNPs 进行了很好的表征。通过荧光光谱和荧光寿命,发现 PNPs 表现出两种类型的发光中心。进一步的机制研究表明,PNPs 的荧光主要来源于三种芳香族氨基酸,即色氨酸、酪氨酸和苯丙氨酸。此外,PNPs 的荧光性质与 pH 值密切相关。PNPs 在恶劣条件和生理条件下都表现出优异的稳定性。此外,PNPs(200μg/mL)对 HeLa 和 GES-1 细胞系没有毒性,表现出良好的生物相容性。只有当细胞受到葡萄糖氧化酶或 HO 胁迫时,PNPs 才会被细胞摄取,随后观察到明亮的蓝色荧光,表明其可用于识别细胞氧化损伤。这些发现将为未来合成甚至更亮的蛋白纳米颗粒及其生物医学应用提供新的线索。

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