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Updated: May 7, 2026

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

The Determination of Protease Specificity in Mouse Tissue Extracts by MALDI-TOF Mass Spectrometry: Manipulating PH to Cause Specificity Changes

Published on: May 25, 2018

Versatile substrates and probes for IgA1 protease activity.

Santosh K Choudary¹, Jiazhou Qiu, Andrew G Plaut

¹Department of Chemistry, Tufts University, 62 Talbot Avenue, Medford MA 02155 (USA).

Chembiochem : a European Journal of Chemical Biology

|September 17, 2013

Summary

Researchers developed novel peptide substrates and a fluorescence probe to study bacterial immunoglobulin A1 (IgA1) proteases. This tool enables quantification of IgA1 protease activity and the screening for inhibitors, aiding infection research.

Keywords:

bacterial meningitis high-throughput screening immunoglobulins peptides proteases

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Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

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Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Preparation and In Vivo Use of an Activity-based Probe for N-acylethanolamine Acid Amidase

Published on: November 23, 2016

Area of Science:

Microbiology
Biochemistry
Enzymology

Background:

Bacterial meningitis is a life-threatening infection.
Pathogenic bacteria like Neisseria, Haemophilus, and Streptococcus use immunoglobulin A1 (IgA1) proteases for colonization and immune evasion.
Existing knowledge of IgA1 protease substrates is limited.

Purpose of the Study:

To design and characterize novel peptide substrates for bacterial IgA1 proteases.
To develop a sensitive assay for quantifying IgA1 protease activity.
To establish a high-throughput screening method for IgA1 protease inhibitors.

Main Methods:

Design and synthesis of diverse peptide substrates based on bacterial IgA1 proteases.
Characterization of protease selectivity profiles using these peptide substrates.
Development of a fluorescence probe assay for IgA1 protease activity measurement.
Application of the probe in buffer and human cerebrospinal fluid.
Establishment of a high-throughput screen for IgA1 protease inhibitors.

Main Results:

Diverse IgA1 proteases exhibited unexpected selectivity profiles on the designed peptide substrates.
A fluorescence probe accurately quantified IgA1 protease activity, detecting Haemophilus influenzae type 1 IgA1 protease at <1 μg/mL.
The first high-throughput screen for IgA1 protease inhibitors was successfully established.

Conclusions:

Novel peptide substrates and a fluorescence probe provide valuable tools for studying bacterial IgA1 proteases.
These tools facilitate the investigation of IgA1 protease roles in bacterial pathogenesis.
The developed screening platform enables the discovery of potential therapeutic inhibitors.