Stereoelectronically Stabilized Flurogenic Probes for Esterase–Activated Biomolecule Imaging

• DOI: 10.1096/fasebj.31.1_supplement.767.18
• Publication Date: 2023-11-26T17:36:45Z
• Authors: Henry R Kilgore; Wen Chyan; Brian Gold; Ronald T Raines

Esterase‐labile fluorogenic probes provide a means of interrogating the spatiotemporal distribution of molecules of interest across biological membranes. In designing such a probe, the primary consideration is maintaining orthogonality of activation to all intracellular reactions except those of interest. We are developing a strategy to curtail ambient hydrolysis of acyl masking groups on fluorescein dyes by carefully balancing n‐to‐pi* and steric interactions to enable the probes to maintain stability in the absence of esterases. The electronic properties of these probes is considered with respect to model compounds that maintain the n‐to‐pi* and steric interactions. These probes were used to visualize and quantify the unusual cytosolic and nuclear transport of large (≥70‐kDa) dextrans. Imaging results with the conjugated probes provide new insight on the permissivity of the nuclear envelope and demonstrate the efficacy of the esterase labile probe strategy.Support or Funding InformationThis work is supported by grant R01 CA073808.