التفاصيل البيبلوغرافية
| العنوان: |
Communication Breakdown: Dissecting the COM Interfaces between the Subunits of Nonribosomal Peptide Synthetases |
| المؤلفون: |
Christopher D. Fage (1339800), Simone Kosol (561429), Matthew Jenner (4067779), Carl Öster (1345275), Angelo Gallo (1718293), Milda Kaniusaite (2551603), Roman Steinbach (11279595), Michael Staniforth (1323402), Vasilios G. Stavros (1263888), Mohamed A. Marahiel (226111), Max J. Cryle (1814797), Józef R. Lewandowski (1279857) |
| سنة النشر: |
2021 |
| المجموعة: |
Smithsonian Institution: Digital Repository |
| مصطلحات موضوعية: |
Biophysics, Biochemistry, Computational Biology, Biological Sciences not elsewhere classified, Chemical Sciences not elsewhere classified, Physical Sciences not elsewhere classified, noncognate domain crosstalk, acceptor C domain, nonribosomal peptide synthetases, carbene footprinting experiments, COM interaction interface, MD, docking domains, bioinformatics analyses show, donor COM region, NMR, COM domains, NRPS, Nonribosomal Peptide Synthetases No. |
| الوصف: |
Nonribosomal peptides are a structurally diverse and bioactive class of natural products constructed by multidomain enzymatic assembly lines known as nonribosomal peptide synthetases (NRPSs). While the core catalytic domains and even entire protein subunits of NRPSs have been structurally elucidated, little biophysical work has been reported on the docking domains that promote interactionsand thus transfer of biosynthetic intermediatesbetween subunits. In the present study, we closely examine the COM domains that mediate COM munication between donor epimerization (E) and acceptor condensation (C) domains found at the termini of NRPS subunits. Through a combination of X-ray crystallography, circular dichroism spectroscopy, solution- and solid-state NMR spectroscopy, and molecular dynamics (MD) simulations, we provide direct evidence for an intrinsically disordered donor COM region that folds into a dynamic helical motif upon binding to a suitable acceptor. Furthermore, our NMR titration and carbene footprinting experiments illuminate the residues involved at the COM interaction interface, and our MD simulations demonstrate folding consistent with experimental data. Although our results lend credence to the previously proposed helix-hand mode of interaction, they also underscore the importance of viewing COM interfaces as dynamic ensembles rather than single rigid structures and suggest that engineering experiments should account for the interactions which transiently guide folding in addition to those which stabilize the final complex. Through activity assays and affinity measurements, we further substantiate the role of the donor COM region in binding the acceptor C domain and implicate this short motif as readily transposable for noncognate domain crosstalk. Finally, our bioinformatics analyses show that COM domains are widespread in natural product pathways and function at interfaces beyond the canonical type described above, setting a high priority for thorough characterization of these docking domains. Our ... |
| نوع الوثيقة: |
dataset |
| اللغة: |
unknown |
| Relation: |
https://figshare.com/articles/media/Communication_Breakdown_Dissecting_the_COM_Interfaces_between_the_Subunits_of_Nonribosomal_Peptide_Synthetases/15175977 |
| DOI: |
10.1021/acscatal.1c02113.s001 |
| الاتاحة: |
https://doi.org/10.1021/acscatal.1c02113.s001 |
| Rights: |
CC BY-NC 4.0 |
| رقم الانضمام: |
edsbas.6E7E6A8D |
| قاعدة البيانات: |
BASE |