Casey Garrett R, Stains Cliff I
Department of Chemistry, University of Nebraska - Lincoln, Lincoln, NE 68588, USA.
Analyst. 2020 Oct 21;145(20):6713-6718. doi: 10.1039/d0an00993h. Epub 2020 Aug 19.
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.
非受体型12磷酸酶（PTPN12或PTP-PEST）是细胞迁移的关键调节因子，在癌症中作为肿瘤抑制因子发挥作用。PTP-PEST表达的降低与肝细胞癌（HCC）的侵袭性表型相关。尽管PTP-PEST在细胞信号传导中很重要，但缺乏直接监测其酶活性的方法。在此，我们报告了一种探针的设计、合成和优化，该探针可通过荧光读数直接监测PTP-PEST的酶活性。这种活性传感器称为pPEST1tide，能够检测低至0.2 nM的重组PTP-PEST。此外，我们证明该探针可以使用一组潜在的脱靶酶选择性地报告PTP-PEST的活性。从长远来看，这种活性探针可用于鉴定PTP-PEST活性的小分子调节剂，并为HCC提供预后读数。
