SENSORS AND ACTUATORS B-CHEMICAL
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2023
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IF 8.4
Ning Xu,Qinglong Qiao,Yi Tao,Pengjun Bao,Yinchan Zhang,Wei Zhou,Jin Li,Zhifeng Li,Zhaochao Xu
Subcellular pH plays a key role to modulate protein structures or specific active sites to impact diverse biological processes. The heterogeneity and high dynamics of intracellular pH require probes that can detect pH in the vicinity of the protein and have a wide pH response range. Here, we reported a protein proximity ligation fluorescent probe for super-resolution imaging of subcellular pH dynamics. Relying on SNAP-tag protein labeling technology, the pH-sensitive fluorophore BG-NDM based on photo-induced electron transfer mechanism was localized to the adjacent positions of the target protein in different subcellular organelles. The equilibrium between the non-fluorescent aggregate and the fluorogenic monomer enabled BG-NDM to have the ability of wash-free super-resolution imaging in live cells. And the wide pH response ranges from pH 6 to pH 12 made the probe have the ability of recognizing pH dynamics. The pH changes around functional proteins during mitophagy, cellular alkalosis and acidosis were revealed. Through long-term super-resolution imaging, we could further record the pH dynamics around mitochondrial proteins COX8A during mitochondrial contacts, fusion and fission at the resolution of a single mitochondria.
研究领域:
生物化学
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细胞生物学
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分子影像学
✓ 已验证
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
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2021
·
IF 10.618
Wenjuan Liu,Qinglong Qiao,Jiazhu Zheng,Jie Chen,Wei Zhou,Ning Xu,Jin Li,Lu Miao,Zhaochao Xu
Super-resolution fluorescence microscopy has emerged as a powerful tool for studying mitochondrial dynamics in living cells. However, the lack of photostable and chemstable probe makes long-term super-resolution imaging of mitochondria still a challenging work. Herein, we reported a 4-azetidinyl-naphthliamide derived SNAP-tag probe AN-BG exhibiting excellent fluorogenicity and photostability for long-term super-resolution imaging of mitochondrial dynamics. The azetidinyl group and naphthalimide fluorophore are in a flat conformation which can effectively suppress twisted intramolecular charge transfer and then effectively improve the brightness and photostability. This planarized molecular structure is conducive to the formation of fluorescence-quenched J -aggregates, and the protein labeling process will depolymerize the probes and restore fluorescence. Fluorescent labeling mitochondrial inner membrane proteins via SNAP tags overcomes the shortcomings that variations in mitochondrial inner membrane potential will release probes attached to mitochondria by electrostatic interactions . Therefore, AN-BG realized the stable labeling of mitochondria and the long-term imaging of mitochondrial dynamics under super-resolution microscopy.
研究领域:
细胞生物学
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分子影像学
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2025
·
IF 3.1
Liju Yu,Jie Gao,Tingting Deng,Chang Gao,Jiaxing Wang,Nan Wang
Heart failure is a global epidemic characterized by high mortality and morbidity rates. β1-Adrenoceptors (ADRβ1) antagonists are the main drugs used in the treatment of HF. A simple and convenient method for screening ADRβ1 antagonists is necessary. Nebivolol has four chiral centers, and its byproducts, which are not easy to avoid, may interact with ADRβ1. Cell membrane chromatography based on SNAP-tag technology provides a comprehensive solution. In this study, an ADRβ1 target cell membrane chromatography model based on SNAP-tag technology was established, and the selectivity, specificity, repeatability, and column life of the system were validated. Under optimized conditions, this model was used to screen for active components targeting ADRβ1 in nebivolol and its byproducts. The bioactivity of nebivolol and its byproducts was detected by calcium influx. The results showed that the ADRβ1 SNAP-tag/CMC model was successfully established and could be used to screen active drugs targeting ADRβ1. Among nebivolol and its byproducts, D-nebivolol (SRRR-enantiomer) had the best affinity for ADRβ1. Three byproducts of nebivolol also exhibited retention characteristics on ADRβ1. In vitro calcium influx experiments verified the interaction of these components with ADRβ1, showing good inhibitory effects on ADRβ1. In conclusion, the ADRβ1 SNAP-tag/CMC model developed in this study can be used for the comprehensive screening of active components targeting ADRβ1 in complex samples.
研究领域:
药理学
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药物筛选
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分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
·
2023
·
IF 3.4
Xin Zhang,Yuhan Jiang,Na Guo,Yifan Ding,Jingting Feng,Chenyang Miao,Yanni Lv
The SNAP-tag-epidermal growth factor receptor (SNAP-tag-EGFR) cell membrane chromatography (CMC) model is a powerful tool for investigating ligand-receptor interactions and screening active ingredients in traditional Chinese medicine. Most tyrosine kinase inhibitors (TKIs) target epidermal growth factor receptors. However, TKIs associated with significant side effects and drug resistance must be addressed immediately. Therefore, there is an urgent need to develop new TKIs with high efficiency and low toxicity. Because of its low toxicity and side effects, traditional Chinese medicine has been widely employed to treat various diseases, including cancer. Hence, this study aimed to use the SNAP-tag-EGFR/CMC-high-performance liquid chromatography-mass spectrometry (HPLC-MS) two-dimensional system model as the research tool to screen and identify potential EGFR antagonists from the Chinese medicine Silybum marianum (L.) Gaertn . The applicability of the system was verified using the positive control drug osimertinib. Four potential EGFR antagonists were screened from the Chinese medicine Silybum marianum (L.) Gaertn. . They were identified as silydianin, silychristin, silybin, and isosilybin. Additionally, their pharmacological activity was preliminarily verified using a CCK-8 assay. The kinetic parameters of the four active ingredients interacting with EGFR and their binding modes with EGFR were analyzed using nonlinear chromatography (NLC) and molecular docking. This study identified silydianin, silychristin, silybin, and isosilybin from Silybum marianum (L.) Gaertn. and verified their potential antitumor effects on EGFR.
研究领域:
药理学
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肿瘤学
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中医药学
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分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）