Ultrasensitive bioluminescent reporters of protein kinase A based on luciferase activity modulated by phosphorylation (PKA LAMP).

ABSTRACT: We developed the novel biosensor platform Luciferase Activity Modulated by Phosphorylation (LAMP) to monitor, with unprecedented sensitivity and dynamic range, reversible protein phosphorylation in cells. Based on NanoLuc luciferase complementation (NanoBiT), LAMP sensors are small (22 kD) and provide stable and bright light output that decreases upon phosphorylation and increases upon dephosphorylation. In this report, we designed LAMP biosensors to report spatial and temporal dynamics of cAMP-dependent protein kinase A (PKA) signaling. By incorporating both PKA phosphorylation and protein phosphatase 2A (PP2A) dephosphorylation motifs into the small component of split NanoLuc (generating PKABiT/pBiT), we achieved a sevenfold dynamic range. LAMP sensors are modular and flexible, allowing

SUBMITTER: Kong Y 

PROVIDER: S-EPMC12329089 | biostudies-literature | 2025 Jul

REPOSITORIES: biostudies-literature