Electrochemical substrate for active profiling of cellular surface leucine aminopeptidase activity and drug resistance in cancer cells
| Autor: | S. Kumaravel, Chun Mao Lin, Yu Chieh Lee, Guo Rong Luo, T.S.T. Balamurugan, Ching Hui Chen, Sheng-Tung Huang, Guan Zhong Chen |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Metallocenes
Biomedical Engineering Biophysics Biosensing Techniques 02 engineering and technology Drug resistance Electrochemistry 01 natural sciences Aminopeptidase Leucyl Aminopeptidase Leucine Limit of Detection Neoplasms medicine Humans Ferrous Compounds Enzyme Assays Cisplatin biology Chemistry digestive oral and skin physiology 010401 analytical chemistry Proteolytic enzymes Electrochemical Techniques Hep G2 Cells General Medicine equipment and supplies 021001 nanoscience & nanotechnology 0104 chemical sciences Biochemistry Drug Resistance Neoplasm Cancer cell biology.protein Antibody 0210 nano-technology human activities Biotechnology medicine.drug |
| Zdroj: | Biosensors and Bioelectronics. 150:111948 |
| ISSN: | 0956-5663 |
| DOI: | 10.1016/j.bios.2019.111948 |
| Popis: | Leucine aminopeptidase (LAP) is an essential proteolytic enzyme and potential biomarker for liver malignancy. Overexpression of LAP is directly linked with some fatal physiological and pathological disorders. In this regard, we have designed an activity based electrochemical substrate leucine-benzyl ferrocene carbamate (Leu-FC) for selective profiling of LAP activity in live cells. In practice, LAP instantaneously hydrolyze the Leu residue of the substrate Leu-FC to eliminate the unmasked electrochemical reporter amino ferrocene via predefined self-immolative cascade. The electrochemical signal is distinctly specific for LAP and free of other electroactive biological interference. The substrate Leu-FC empowered sensor displayed broad dynamic range with admirable detection limits. On top of this, the probe Leu-FC was employed in real-time active profiling of cellular LAP activity in HepG2 cells and effect of LAP inhibitor. In extent, the substrate Leu-FC can effectively monitor cisplatin induced overexpression of LAP activity in HepG2 cells in presence and absence of bestatin. The sensor showcased an excellent reliability towards monitoring cellular LAP activity in HepG2 cells. Unlike the traditional antibody-based immunoassays, our approach is capable of monitoring in-situ activity of LAP in live cells. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect