A fluorescent probe for monitoring PTP-PEST enzymatic activity. Issue 20 (19th August 2020)

• Record Type: Journal Article
• Authors: Casey, Garrett R.; Stains, Cliff I.
• Is Part Of: Analyst. Volume 145:Issue 20(2020)
• Page Start: 6713
• Page End: 6718
• Publication Date: 2020-08-19
• Subjects: Chemistry, Analytic -- Periodicals; 543
• DOI: 10.1039/d0an00993h
• Languages: English
• ISSNs: 0003-2654

Abstract

Abstract : Design and optimization of a direct activity assay for PTP-PEST. Abstract : Phosphatase non-receptor type 12 (PTPN12 or PTP-PEST) is a critical regulator of cell migration, acting as a tumor suppressor in cancer. Decreases in PTP-PEST expression correlate with aggressive phenotypes in hepatocellular carcinoma (HCC). Despite the importance of PTP-PEST in cellular signaling, methods to directly monitor its enzymatic activity are lacking. Herein, we report the design, synthesis, and optimization of a probe to directly monitor PTP-PEST enzymatic activity via a fluorescent readout. This activity sensor, termed pPEST1tide, is capable of detecting as little as 0.2 nM recombinant PTP-PEST. In addition, we demonstrate that this probe can selectively report on PTP-PEST activity using a panel of potential off-target enzymes. In the long-term, this activity probe could be utilized to identify small molecule modulators of PTP-PEST activity as well as provide a prognostic readout for HCC.