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Constructing Cyclic Peptides Using an On-Tether Sulfonium Center
Published on: September 28, 2022
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Crosslinking intermodular condensation in non-ribosomal peptide biosynthesis.
Graham W Heberlig1, James J La Clair1, Michael D Burkart2
1Department of Chemistry and Biochemistry, University of California San Diego, La Jolla, CA, USA.
Nature
|December 11, 2024
Summary
Researchers developed novel crosslinking probes to study non-ribosomal peptide synthetases (NRPS). These probes captured key interactions, revealing the structural dynamics of these complex enzyme assembly lines for therapeutic drug production.
Area of Science:
- Biochemistry
- Structural Biology
- Molecular Biology
Background:
- Non-ribosomal peptide synthetases (NRPS) are large megasynthase enzymes crucial for producing therapeutic drugs.
- Their complex, dynamic, multi-domain architecture poses significant challenges for structural characterization.
- Understanding NRPS function requires detailed insights into inter-domain and inter-modular interactions.
Purpose of the Study:
- To develop and apply novel crosslinking probes for conformational constraint of NRPS.
- To resolve the dynamic interactions between peptidyl carrier proteins and enzymatic domains within NRPS.
- To elucidate the structural basis of substrate translocation and catalytic progression in NRPS.
Main Methods:
- Development of site-selective crosslinking probes.
- Application of tetrazine click chemistry for in situ crosslinking.
- High-resolution cryo-electron microscopy (cryo-EM) and X-ray crystallography.
Main Results:
- Captured the condensation of two carrier protein substrates within the tyrocidine synthetase active site.
- Determined the high-resolution cryo-EM structure of this crosslinked complex.
- Obtained X-ray crystal structure of a carrier protein crosslinked to its epimerization domain.
- Highlighted intermodular recognition events and defined carrier protein processive movement.
Conclusions:
- Structural insights into NRPS intermodular recognition and carrier protein dynamics were achieved.
- The study provides a framework for understanding the molecular mechanisms of NRPS assembly lines.
- These findings are critical for future synthetic biology applications and rational design of NRPS-based pathways.
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