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Dual colorimetric and fluorogenic probes for visualizing tyrosine phosphatase activity.

Suvendu Biswas¹, Brandon S McCullough¹, Elena S Ma¹

¹Department of Medicinal Chemistry, University of Utah College of Pharmacy, Salt Lake City, Utah 84112, USA. amy.barrios@utah.edu.

Chemical Communications (Cambridge, England)

|February 2, 2017

Summary

New resorufin-based substrates offer sensitive detection of protein tyrosine phosphatase (PTP) activity in vitro and in cells. These tools also enable visual detection of Staphylococcus aureus using a colorimetric readout.

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Area of Science:

Biochemistry
Analytical Chemistry
Microbiology

Background:

Protein tyrosine phosphatases (PTPs) are crucial enzymes involved in cellular signaling.
Dysregulation of PTP activity is linked to various diseases, necessitating robust detection methods.
Existing methods for PTP activity detection can be limited in sensitivity or applicability.

Purpose of the Study:

To synthesize novel resorufin-based substrates for PTP activity.
To evaluate the substrates' performance for fluorogenic detection of PTPs in vitro and in living cells.
To explore the utility of these substrates for detecting pathogenic bacteria.

Main Methods:

Chemical synthesis of two resorufin-based fluorogenic substrates.
Enzyme assays to measure PTP activity using the synthesized substrates.
Cell-based assays to assess PTP activity in living cells at different pH levels.
Colorimetric detection assays for bacterial presence.

Main Results:

Successfully synthesized two novel resorufin-based substrates.
Demonstrated sensitive fluorogenic readout of PTP activity across acidic and neutral pH.
Achieved sensitive detection of PTP activity in vitro and within living cells.
Visual colorimetric detection of Staphylococcus aureus was achieved.

Conclusions:

The developed resorufin-based substrates are versatile tools for sensitive PTP activity monitoring.
These substrates are applicable in diverse biological settings, including cell-based assays and bacterial detection.
The findings offer new avenues for PTP research and diagnostics.