利用配体引导的荧光探针绘制活细胞中的β3-肾上腺素能受体图谱

IF 6.7 1区 化学 Q1 CHEMISTRY, ANALYTICAL Analytical Chemistry Pub Date : 2024-10-30 DOI:10.1021/acs.analchem.4c03408
Yani Liu, Xiangjie Luo, Wei Cheng, Ling Zhou, Yang Zhou, Hai-Liang Zhu, Tony D. James, Yong Qian
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引用次数: 0

摘要

实时跟踪活细胞或组织中原生蛋白质的动态分布对于了解其生理或病理过程中的功能机制至关重要。β3-肾上腺素能受体(β3-AR)具有重要的生物学功能。它有望成为衰老的诊断指标,但目前的β-AR探针无法实时动态监测β3-AR,这阻碍了β3-AR研究的进展。在这里,我们开发了一种配体定向锚定探针--β3-ARP,它能在活细胞中与原生β3-AR精确共价锚定,用于衰老诊断。这种配体定向锚定探针可选择性地识别和追踪β3-AR,实现对活细胞中β3-AR的动态观察和监测,包括脂滴和线粒体之间的相互作用。此外,我们还能直接原位探测衰老小鼠各器官中β3-AR的分布,并利用β3-ARP和双光子成像技术追踪不同类型原代神经元细胞中原生β3-AR的位置,揭示了神经系统疾病中脂滴的异常堆积和β3-AR的分布。这项研究产生了一种新的共价锚定荧光探针--β3-ARP,它可以作为探索β3-AR与老年相关疾病之间联系的有力工具。
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Mapping of β3-Adrenergic Receptor in Living Cells with a Ligand-Guided Fluorescent Probe
Tracking the dynamic distribution of native proteins in living cells or tissues in real time is essential for understanding the functional mechanisms involved in their physiological or pathological processes. The β3-adrenergic receptor (β3-AR) has important biological functions. It is expected to be a diagnostic indicator for aging, yet current probes for β-ARs are unable to dynamically monitor β3-AR in real time, which impedes the progress of β3-AR research. Here, we developed a ligand-directed anchoring probe, β3-ARP, that precisely covalently anchors with native β3-AR in living cells, for the diagnosis of senescence. The ligand-directed anchoring probe selectively recognizes and traces β3-AR and can achieve dynamic observation and monitoring of β3-AR in living cells, including the interaction between lipid droplets and mitochondria. Moreover, we were able to directly probe the distribution of β3-AR in various organs of aging mice in situ and track the location of native β3-AR in different types of primary neuronal cells using β3-ARP and two-photon imaging, which revealed the aberrant accumulation of lipid droplets and the distribution of β3-AR in neurological diseases. This research has resulted in a new covalently anchoring fluorescent probe, β3-ARP, which could serve as a powerful tool to explore the link between β3-AR and age-related diseases.
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来源期刊
Analytical Chemistry
Analytical Chemistry 化学-分析化学
CiteScore
12.10
自引率
12.20%
发文量
1949
审稿时长
1.4 months
期刊介绍: Analytical Chemistry, a peer-reviewed research journal, focuses on disseminating new and original knowledge across all branches of analytical chemistry. Fundamental articles may explore general principles of chemical measurement science and need not directly address existing or potential analytical methodology. They can be entirely theoretical or report experimental results. Contributions may cover various phases of analytical operations, including sampling, bioanalysis, electrochemistry, mass spectrometry, microscale and nanoscale systems, environmental analysis, separations, spectroscopy, chemical reactions and selectivity, instrumentation, imaging, surface analysis, and data processing. Papers discussing known analytical methods should present a significant, original application of the method, a notable improvement, or results on an important analyte.
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