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Specificity profiling of human trypsin-isoenzymes
Oliver Schilling1,2,3, Martin L Biniossek1, Bettina Mayer1
1Institute of Molecular Medicine and Cell Research, Faculty of Medicine, University of Freiburg, D-79104 Freiburg, Germany.
Biological Chemistry
|June 9, 2018
Summary
Researchers studied human trypsin isoenzymes, finding distinct cleavage specificities. Trypsin-3 shows unique preferences, potentially aiding in developing targeted cancer therapies and diagnostic tools.
Area of Science:
- Biochemistry
- Proteomics
- Enzymology
Background:
- Humans possess three trypsin isoenzymes, with trypsin-3 exhibiting unique functional characteristics.
- Understanding trypsin isoenzyme specificity is crucial for biochemical research and therapeutic development.
Purpose of the Study:
- To systematically investigate and compare the substrate specificity of human trypsin isoenzymes (trypsin-1, -2, and -3).
- To elucidate the molecular basis for trypsin-3's unique catalytic activity and its potential as a therapeutic target.
Main Methods:
- Utilized proteome-derived peptide libraries and quantitative proteomics to analyze cleavage site preferences.
- Employed Förster Resonance Energy Transfer (FRET) peptide substrates to validate specificity findings.
- Performed structural analysis to explain trypsin-3's unique substrate interactions.
Main Results:
- All three trypsin isoenzymes exhibit a primary preference for cleavage after lysine and arginine residues.
- Trypsin-1 shows a slight preference for lysine, while trypsin-3 does not discriminate between lysine and arginine.
- Trypsin-3 displays unique P1' and P2' preferences for threonine and aspartic acid, respectively, explained by a salt bridge interaction with Arg193.
Conclusions:
- The study reveals distinct substrate specificity profiles for human trypsin isoenzymes, particularly highlighting trypsin-3's unique characteristics.
- The identified specificity of trypsin-3, driven by a salt bridge mechanism, offers insights for developing selective inhibitors.
- Findings support the potential of trypsin-3 as a therapeutic target and diagnostic marker for cancers, aiding in the development of novel cancer therapies.
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