Proteomic profiling of proteases: tools for granzyme degradomics

Area of Science:

• Biochemistry
• Proteomics
• Enzymology

Background:

• Protease dysfunction is implicated in numerous human diseases.
• Despite encoding 2% of the genome, few proteases have well-defined functions.
• Identifying protease specificity and substrates in biological samples remains challenging.

Purpose of the Study:

• To summarize and discuss current proteomic techniques for identifying protease specificity.
• To highlight methods for determining protease cleavage sites and natural substrates.
• To focus on cytotoxic lymphocyte granule serine proteases (granzymes).

Main Methods:

• Proteomic screens are rapidly advancing.
• Techniques include peptide libraries, mass spectrometry-based methods (2D-DIGE, differential isotope labeling), quantitative degradomics, and activity-based protein profiling.
• These methods aid in identifying protease activity and targets.

Main Results:

• Impressive progress has been made in proteomic screens for protease identification.
• Various techniques allow for the characterization of protease specificity and cleavage sites.
• The review consolidates current knowledge on these advanced methods.

Conclusions:

• Proteomic approaches are essential for elucidating protease functions.
• Understanding protease specificity is key to developing therapeutics for protease-related diseases.
• Further application of these techniques, especially for granzymes, will advance biological and medical research.