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基于拉曼光谱的线粒体介导凋亡过程中分子特征变化的原位监测

Raman-Based in Situ Monitoring of Changes in Molecular Signatures during Mitochondrially Mediated Apoptosis.

作者信息

Shin Hyeon Jeong, Lee Ji Hye, Kim Yong Duk, Shin Injae, Sim Taebo, Lim Dong-Kwon

机构信息

KU-KIST Graduate School of Converging Science and Technology, Korea University, 145 Anam-ro, Seongbuk-gu, Seoul 02841, Republic of Korea.

Chemical Kinomics Research Center, Korea Institute of Science and Technology, 5 Hwarangro 14-gil, Seongbuk-gu, Seoul, South Korea.

出版信息

ACS Omega. 2019 May 6;4(5):8188-8195. doi: 10.1021/acsomega.9b00629. eCollection 2019 May 31.

Abstract

Obtaining molecular information from inside cells is an important topic to understand the outcome of molecular interactions between potential drug molecules and biomolecules inside cells. To envision this goal, we investigated the surface-enhanced Raman scattering-based single-cell spectroscopic method to monitor changes in intracellular molecular signatures during mitochondrially mediated apoptosis in real time. Triphenylphosphine-modified gold nanoparticles were localized successfully to the mitochondria and greatly enhanced to obtain the intrinsic Raman scattering spectrum of mitochondria and cytochrome in the live cell. Photothermally induced apoptosis showed a moderate decrease in the disulfide bond and a sharp increase in β-sheet structures depending on the input-laser power, along with morphological changes. However, chemical drug induced-apoptosis showed more subtle changes in the disulfide bond, as well as changes in Raman peaks corresponding to cytochrome , and the appearance of a new peak at 1420 cm, which enabled us to study the molecular interactions within the mitochondria in real time from a single cell, following treatment with a novel pyruvate dehydrogenase kinase inhibitor.

摘要

从细胞内部获取分子信息是理解潜在药物分子与细胞内生物分子之间分子相互作用结果的一个重要课题。为了实现这一目标,我们研究了基于表面增强拉曼散射的单细胞光谱方法,以实时监测线粒体介导的细胞凋亡过程中细胞内分子特征的变化。三苯基膦修饰的金纳米颗粒成功定位于线粒体,并得到极大增强,从而获得活细胞中线粒体和细胞色素的固有拉曼散射光谱。光热诱导的细胞凋亡显示,根据输入激光功率的不同,二硫键适度减少,β-折叠结构急剧增加,同时伴有形态变化。然而,化学药物诱导的细胞凋亡在二硫键上显示出更细微的变化,以及与细胞色素相对应的拉曼峰的变化,并且在1420 cm处出现了一个新峰,这使我们能够在用新型丙酮酸脱氢酶激酶抑制剂处理后,从单个细胞实时研究线粒体内的分子相互作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4544/6648662/01a82e6eb7e6/ao-2019-006293_0001.jpg

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