Precipitated Fluorophore-Based Molecular Probe for In Situ Imaging of Aminopeptidase N in Living Cells and Tumors
| Autor: | Yongchao Liu, Chengyan Xu, Hong-Wen Liu, Lili Teng, Lin Yuan, Shuangyan Huan, Xiao-Bing Zhang |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Alanine
chemistry.chemical_classification In situ animal structures Fluorophore Endoplasmic reticulum 010401 analytical chemistry 010402 general chemistry 01 natural sciences Fluorescence 0104 chemical sciences Analytical Chemistry Amino acid chemistry.chemical_compound chemistry Cytoplasm Biophysics Molecular probe hormones hormone substitutes and hormone antagonists |
| Zdroj: | Analytical Chemistry. 93:6463-6471 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.1c00280 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|