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Updated: Jul 17, 2026
06:53
In Vivo Monitoring of Transcriptional Activity During Metabolic Transition Using a Bioluminescent Reporter in Yeast
Published on: February 21, 2025
A self-immolative reporter for beta-galactosidase sensing.
Nan-Hui Ho1, Ralph Weissleder, Ching-Hsuan Tung
1Center of Molecular Imaging Research, Massachusetts General Hospital, Harvard Medical School, 149 13th Street, Room 5406, Charlestown, MA 02129, USA.
Chembiochem : a European Journal of Chemical Biology
|February 16, 2007
Summary
A new dual probe, Gal-2SBPO, enables sensitive detection of beta-galactosidase activity. This probe releases a detectable signal upon enzymatic cleavage, offering a valuable tool for biochemical assays.
Area of Science:
- Biochemistry
- Chemical Biology
- Molecular Probes
Background:
- Beta-galactosidase is a key enzyme with implications in various biological processes and diseases.
- Development of sensitive and specific probes is crucial for accurate enzyme activity assays.
- Existing probes may lack dual detection capabilities or optimal stability under physiological conditions.
Purpose of the Study:
- To develop a novel dual fluorogenic and chromogenic probe for beta-galactosidase.
- To synthesize and characterize the probe, Gal-2SBPO, for its stability and solubility.
- To validate the probe's utility in detecting beta-galactosidase activity.
Main Methods:
- Synthesis of a water-soluble dye, 9-di-3-sulfonyl-propylaminobenzo[a]phenoxazonium perchloride (2SPBO).
- Conjugation of beta-D-galactopyranoside to 2SPBO via a self-immolative spacer and molecular blocker.
- Assay of beta-galactosidase activity by monitoring absorbance (630 nm) and fluorescence (670 nm) changes.
Main Results:
- The developed probe, Gal-2SBPO, demonstrated good solubility and stability under physiological conditions.
- Enzymatic cleavage by beta-galactosidase triggered a cascade leading to the release of the active 2SPBO dye.
- The probe allowed for sensitive detection of beta-galactosidase activity through dual optical readouts.
Conclusions:
- Gal-2SBPO is an effective dual fluorogenic and chromogenic probe for beta-galactosidase.
- The probe's design facilitates a clear signal upon enzymatic activity.
- This probe offers a versatile tool for biochemical research and diagnostics involving beta-galactosidase.
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