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Bacterial Peptide Display for the Selection of Novel Biotinylating Enzymes
Published on: October 3, 2019
In vitro selection of functional lantipeptides.
Frank T Hofmann1, Jack W Szostak, Florian P Seebeck
1Department of Physical Biochemistry, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany.
Journal of the American Chemical Society
|May 2, 2012
Summary
Researchers developed a novel method to create and screen billions of artificial lantipeptides for function. This approach successfully identified lantipeptides that bind to Sortase A, a key virulence factor in Staphylococcus aureus.
Area of Science:
- Biochemistry
- Molecular Biology
- Synthetic Biology
Background:
- Lantipeptides are ribosomally synthesized peptides characterized by the presence of lanthionine cross-links.
- Identifying functional lantipeptides with specific biological activities is challenging due to their structural complexity and diversity.
- Sortase A is a crucial transamidase enzyme for the pathogenesis of Staphylococcus aureus, making it an attractive therapeutic target.
Purpose of the Study:
- To develop a high-throughput method for generating and identifying functional artificial lantipeptides.
- To validate the utility of this method by isolating novel binders to Sortase A.
- To investigate the structural requirements for lantipeptide interaction with Sortase A.
Main Methods:
- Utilized in vitro translation coupled with an enzyme-free posttranslational modification protocol to generate a diverse library of over 10^11 artificial lantipeptides.
- Employed mRNA-lantipeptide display for high-throughput screening of the lantipeptide library.
- Biochemical assays were used to characterize the interaction between isolated lantipeptides and Sortase A.
Main Results:
- Successfully generated a vast library of lanthionine-containing peptides with unprecedented diversity.
- Identified specific lantipeptide binders that exhibit high affinity for Sortase A.
- Demonstrated that the binding interaction is critically dependent on the presence of a (2S,6R)-lanthionine residue and the active conformation of Sortase A.
Conclusions:
- The developed mRNA-lantipeptide display method is effective for discovering functional peptides from highly diverse libraries.
- This approach provides a powerful tool for identifying novel inhibitors or modulators of enzymes like Sortase A.
- Understanding the structure-activity relationship is crucial for designing potent lantipeptide-based therapeutics against bacterial infections.
