An Integrated Chemical Cytometry Method: Shining a Light on Akt Activity in Single Cells. Issue 42 (6th October 2016)

• Record Type: Journal Article
• Authors: Mainz, Emilie R.; Wang, Qunzhao; Lawrence, David S.; Allbritton, Nancy L.
• Is Part Of: Angewandte Chemie international edition. Volume 55:Issue 42(2016)
• Page Start: 13095
• Page End: 13098
• Publication Date: 2016-10-06
• Subjects: biosensors -- chemical cytometry -- kinase activity probes -- photochemistry -- single-cell analysis; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201606914
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: 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. Abstract : See the light : An integrated analysis system that utilizes a light‐programmable, cell‐permeable reporter that can be delivered simultaneously into many cells was developed. The reporter is a substrate for Akt, a key oncogenic kinase, and was used to evaluate the Akt activity of statistically significant numbers of primary human cells through automated single‐cell capillary electrophoresis.