期刊
ANALYTICAL CHEMISTRY
卷 93, 期 16, 页码 6463-6471出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c00280
关键词
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资金
- National Natural Science Foundation of China [21977027, 21890744, 21877029]
- National Key R&D Program of China [2019YFA0210100]
- National Postdoctoral Program for Innovative Talents [BX20180093]
- Hunan Postgraduate Research and Innovation Project [CX2018B187]
This study introduced a novel precipitated fluorophore HBPQ which is completely insoluble in water and can be controlled for solid fluorescence through protection-deprotection of phenolic hydroxyl. A APN-responsive fluorogenic probe (HBPQ-A) was successfully designed using HBPQ for imaging intracellular APN with stable and reliable fluorescent signals.
Aminopeptidase N (APN) is capable of cleaving N-terminal amino acids from peptides with alanine in the N-terminal position and plays a key role in the growth, migration, and metastasis of cancer. However, reliable in situ information is hard to be obtained with the current APN-responsive molecular probes because the released fluorophores are cytoplasmic soluble and thus rapidly depart from the enzymatic reaction sites and spread out all over the cytoplasm. Here, we report a de novo precipitated fluorophore, HBPQ, which is completely insoluble in water and shows strong yellow solid emission when excited with a 405 nm laser. Owing to the controllable solid fluorescence of HBPQ by the protection-deprotection of phenolic hydroxyl, we further utilized HBPQ to design an APN-responsive fluorogenic probe (HBPQ-A) for the imaging of intracellular APN. Importantly, HBPQ-A can not only perform in situ imaging of APN in different organelles (e.g., lysosomes, mitochondria, endoplasmic reticula, and so forth) but also display a stable and indiffusible fluorescent signal for reliable mapping of the distribution of APN in living cells. In addition, through real-time imaging of APN in 4T1 tumors, we found that the fluorescent signal with high fidelity generated by HBPQ-A could remain constant even after 12 h, which further confirmed its diffusion-resistant ability and long-term reliable imaging ability. We believe that the precipitated fluorophore may have great potential for long-term in situ imaging.
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