An Integrated Chemical Cytometry Method: Shining a Light on Akt Activity in Single Cells
| Autor: | Qunzhao Wang, David S. Lawrence, Emilie R. Mainz, Nancy L. Allbritton |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Chemistry Kinase 010401 analytical chemistry General Chemistry Biosensing Techniques Flow Cytometry 01 natural sciences Catalysis Article 0104 chemical sciences 03 medical and health sciences 030104 developmental biology Biochemistry Single-cell analysis Cytoplasm Cell Line Tumor Ultraviolet light Moiety Humans Kinase activity Single-Cell Analysis Cytometry Protein kinase B Proto-Oncogene Proteins c-akt |
| Zdroj: | Angewandte Chemie (International ed. in English). 55(42) |
| ISSN: | 1521-3773 |
| Popis: | Tools to evaluate oncogenic kinase activity in small clinical samples have the power to guide precision medicine in oncology. Existing platforms have demonstrated impressive insights into the activity of protein kinases, but these technologies are unsuitable for the study of kinase behavior in large numbers of primary human cells. To address these limitations, we developed an integrated analysis system that utilizes a light-programmable, cell-permeable reporter deliverable simultaneously to many cells. The reporter's ability to act as a substrate for Akt, a key oncogenic kinase, was masked by a 2-4,5-dimethoxy 2-nitrobenzyl (DMNB) moiety. Upon exposure to ultraviolet light and release of the masking moiety, the substrate sequence enabled programmable reaction times within the cell cytoplasm. When coupled to automated single-cell capillary electrophoresis, statistically significant numbers of primary human cells were readily evaluated for Akt activity. |
| Databáze: | OpenAIRE |
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