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Cell-Membrane-Localizing Fluorescence Probes for Aminopeptidase N
Yun Jae Yang1, Mingchong Dai2, Kyo Han Ahn1
1Department of Chemistry, Pohang University of Science and Technology, Pohang 37673, South Korea.
ACS Sensors
|July 5, 2023
Summary
New probes detect Aminopeptidase N (APN) activity on cell membranes, overcoming limitations of internal probes. This enables accurate APN level measurement in tissues and disease models, aiding diagnosis and research.
Area of Science:
- Biochemistry
- Molecular Biology
- Medical Imaging
Background:
- Aminopeptidase N (APN) is a transmembrane enzyme involved in cell functions and disease.
- Elevated APN levels are observed in tumors and damaged organs, necessitating noninvasive detection methods.
- Existing APN probes measure intracellular fluorescence, leading to inaccurate data due to cell permeability and enzyme kinetics.
Purpose of the Study:
- To develop novel cell-membrane-localizing probes for accurate APN activity detection.
- To address the limitations of current intracellular probes for APN monitoring.
- To enable reliable quantification of APN levels in biological tissues and disease models.
Main Methods:
- Development of two small-molecule probes that localize to the cell membrane.
- Probes generate ratiometric fluorescence signals upon APN enzymatic activity.
- Utilized two-photon imaging for APN level determination in various organs and disease models.
Main Results:
- Developed probes selectively respond to APN with ratiometric fluorescence changes.
- Quantified relative APN levels in mouse organs: intestine (4.3), kidney (2.1), liver (2.7), lung (3.2), and stomach (1.0).
- Observed elevated APN levels in HepG2-xenograft tumors and acetaminophen-induced liver injury models.
Conclusions:
- The novel probes provide a reliable method for studying APN-associated biology.
- Enables accurate, noninvasive assessment of APN levels in tissues and disease states.
- Facilitates research into APN's role in diseases like drug-induced hepatotoxicity.
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