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Capturing Covalent Catalytic Intermediates by Enzyme Mutants: Recent Advances in Methodologies and Applications
1The First Affiliated Hospital of USTC, Centre for Advanced Interdisciplinary Science and Biomedicine of IHM, MOE key Laboratory for Cellular Dynamics, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, 230001, P. R. China.
Chembiochem : a European Journal of Chemical Biology
|March 3, 2023
Summary
Researchers developed chemical strategies to stabilize transient covalent intermediates for studying enzyme mechanisms. Enzyme mutants, like those with 2,3-diaminopropionic acid, are key for trapping these crucial enzyme-substrate interactions.
Area of Science:
- Biochemistry
- Enzymology
- Chemical Biology
Background:
- Covalent catalytic intermediates are vital for understanding enzyme mechanisms, activity, and substrate specificity.
- Naturally occurring intermediates are often too unstable for detailed study.
- Chemical strategies are needed to stabilize these transient species for analysis.
Purpose of the Study:
- To review mechanism-based strategies for trapping covalent catalytic intermediates.
- To highlight enzyme mutant approaches, particularly using 2,3-diaminopropionic acid.
- To discuss applications and future directions of enzyme substrate traps.
Main Methods:
- Summarization of three general mechanism-based strategies for trapping covalent intermediates.
- Detailed description of enzyme mutant-based approaches, including genetic encoding of 2,3-diaminopropionic acid.
- Presentation of applications in structural, functional, and protein labeling studies.
Main Results:
- Established strategies for stabilizing enzyme-substrate intermediates.
- Demonstrated the utility of specific enzyme mutants for trapping acyl-enzyme intermediates in proteases.
- Showcased applications of trapped intermediates in various biological studies.
Conclusions:
- Enzyme substrate traps are essential tools for investigating catalytic mechanisms.
- Mutant-based strategies offer powerful methods for capturing transient intermediates.
- Further development of enzyme substrate traps holds promise for future biochemical research.
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